Effect of Clomiphene Citrate on the Ultrastructure of Testis in Albino Rats

SUMMARY: The aim of the present work was to study the closer effect of clomiphene citrate on the ultrastructure of the testis of adult albino rats to provide a basis for optimizing this drug in the treatment of male infertility. The testes were removed from both groups under anesthesia and then prepared for examination by light using hematoxylin and eosin stains and a transmission electron microscope. Semithin sections were cut into 1 µm thick sections, stained with toluidine blue, and examined by light microscopy for a survey. The desired areas were placed in the center, and other areas were trimmed. Primary spermatocytes showed marked nuclear changes (pyknosis), and their nuclear membranes were ill-defined and disrupted. The cytoplasm showed widespread degeneration of mitochondria and lysosomes and focal degeneration of the rough endoplasmic reticulum compared with the control group. The spermatids were pale, and the two phases of spermatogenesis were distinctly identifiable in the control group but were confused in the treated group. Some spermatids had interrupted nuclear membranes, also containing degenerated mitochondria, focal fragmentation of rough endoplasmic reticulum, and free ribosomes. Spermatozoa in the treated group appeared deformed compared to the control, where they had deformed head caps. Leydig cells of the treated group have an irregularly shaped nucleus, with focal chromatin aggregation and peripheral chromatin condensation on the inner surface of the nuclear membrane. The observations of the present work indicate a possible causal relationship between testicular affection and ingestion of clomiphene citrate, which can be avoided by close medical observations using ultrasonography, semen analysis, or testicular biopsy to detect early malignant changes. Furthermore, the drug should not be used for more than three to six cycles and should be stopped for at least three cycles before reuse. When clomiphene citrate is ineffective in the treatment of male infertility, human menopausal gonadotropin (hMG) administration is typically selected. However, high-dose hMG therapy is associated with a variety of adverse effects. In this work, we report the success of a modified clomiphene citrate regimen in increasing sperm count without any hazards to the testicular tissue.

El objetivo del trabajo fue estudiar el efecto del citrato de clomifeno sobre la estructura de los testículos de la rata albina adulta, con la finalidad de determinar la mejor manera de utilizar este fármaco en el tratamiento de la infertilidad masculina. Los testículos se extrajeron bajo anestesia y para su análisis a través de microscopio de luz se tiñeron con HE. Además, las muestras fueron preparadas para su examen con microscopía electrónica de transmisión. Por otra parte, se cortaron secciones semifinas de 1 µm de espesor, se tiñeron con azul de toluidina y se examinaron mediante microscopía óptica. Los espermatocitos primarios mostraron cambios nucleares marcados (picnosis) y sus membranas nucleares estaban mal definidas y alteradas. En el grupo experimental las células presentaban el citoplasma con degeneración generalizada de las mitocondrias y de los lisosomas y una degeneración focal del retículo endoplásmico rugoso en comparación con el grupo control. Las espermátidas estaban pálidas y las dos fases de la espermatogénesis eran claramente identificables en el grupo control, pero se confundían en el grupo tratado. Algunas espermátidas tenían membranas nucleares interrumpidas, y también contenían mitocondrias degeneradas, fragmentación focal del retículo endoplásmico rugoso y ribosomas libres. Los espermatozoides del grupo tratado se presentaban deformados en comparación con el control. Las células de Leydig del grupo tratado presentaban un núcleo de forma irregular, con agregación focal de cromatina y condensación de cromatina periférica en la superficie interna de la membrana nuclear. Las observaciones del presente trabajo indican una posible relación causal entre la afección testicular y la ingestión de citrato de clomifeno, que puede evitarse mediante observaciones médicas minuciosas a través de ecografía, análisis de semen o biopsia testicular para detectar cambios malignos tempranos. Además, el medicamento no debiera ser usado durante más de tres a seis ciclos y debe suspenderse durante al menos tres ciclos antes de volver a usarlo. Cuando el citrato de clomifeno es ineficaz en el tratamiento de la infertilidad masculina, normalmente se selecciona la administración de gonadotropina menopáusica humana (hMG). Sin embargo, la terapia con hMG en dosis altas se asocia con una variedad de efectos adversos. En este trabajo, informamos el éxito de un régimen modificado con citrato de clomifeno para aumentar el recuento de espermatozoides sin riesgo para el tejido testicular.

Gamma-amino butyric acid immunoreactivity in the testis tissue of Wistar and genetic absence epilepsy rats / Inmunorreactividad del ácido gamma-amino butírico en el tejido testicular de ratas Wistar con epilepsia de ausencia genética

SUMMARY: Studies have shown the adverse effects of epileptic seizures on reproductive health. The aim of the present study was to investigate morphological changes, apoptosis and GABA localization in the testis tissue of genetic absence epilepsy rats. Testis tissues of GAERS and Wistar rats were processed for paraffin embedding and electron microscopy. Sections were stained with hematoxylin and eosin, Masson's trichrome and periodic acid-Schiff reaction. GABA immunohistochemistry was applied for determining the alterations in GABA levels. GABA immunoreactivity was observed in the seminiferous tubules and interstitial areas of both GAERS and Wistar rats. GABA immunoreactivity was found to be decreased in GAERS compared to Wistar group. Electron microscopic observations showed that GABA was present in manchette microtubules, sperm tail and neck at different phases of spermiogenesis. Qualitative observations revealed that testis tissues of GAERS showed reduced sperm in the seminiferous tubules compared to the Wistar controls. In conclusion, we demonstrated GABAergic system in the seminiferous tubules of control and GAERS rats, in parallel with the previous studies; and there were alterations in this system in GAERS. We suggest that these alterations in absence epilepsy may also affect the gonadal system, resulting in decreased sperm production.

RESUMEN: Los estudios han demostrado los efectos adversos de las convulsiones epilépticas sobre la salud reproductiva. El objetivo del presente estudio fue investigar los cambios morfológicos, la apoptosis y la localización de GABA en el tejido testicular de ratas con epilepsia de ausencia genética. Se procesaron tejidos testiculares de ratas GAERS y Wistar para inclusión en parafina y microscopía electrónica. Las secciones se tiñeron con hematoxilina y eosina, tricrómico de Masson y reacción de ácido peryódico de Schiff. Se aplicó inmunohistoquímica de GABA para determinar las alteraciones en los niveles de GABA. Se observó inmunorreactividad de GABA en los túbulos seminíferos y las áreas intersticiales de las ratas GAERS y Wistar. Se encontró que la inmunorreactividad de GABA estaba disminuida en GAERS en comparación con el grupo Wistar. Las observaciones microscópicas electrónicas mostraron que GABA estaba presente en los microtúbulos, la cola y el cuello del espermatozoide en diferentes fases de la espermiogénesis. Las observaciones cualitativas revelaron que los tejidos testiculares de GAERS mostraron una reducción de los espermatozoides en los túbulos seminíferos en comparación con los controles Wistar. En conclusión, demostramos el sistema GABAérgico en los túbulos seminíferos de ratas control y GAERS, en paralelo con estudios previos; y además se observaron alteraciones en este sistema en GAERS. Sugerimos que estas alteraciones en epilepisa de ausencia genética también pueden afectar el sistema gonadal, resultando en una disminución de la producción de semen.

Potential radioprotective effect of AT1 receptor antagonists against morphological and ultrastructural changes in the testes induced by ionizing radiation / Potencial efecto radioprotector de antagonistas del receptor AT1 contra los daños morfológicos y ultraestructurales en los testículos inducido por la radiación ionizante

Radiotherapy is a source of human exposure to ionizing radiation. This pure energy causes deleterious effects on tissues, which result from oxidative stress, a phenomenon in which there is the participation of the Renin-Angiotensin System (RAS). The male genital organs are extremely radiosensitive and the action of radiation in the testes can significantly affect spermatogenesis. In search of potential radioprotective for male genital system, this study investigated whether the AT1 receptor antagonists minimize radiation-induced damage to reproductive tissues, by decreasing oxidative stress. Male Wistar rats were divided into six groups: 0 Gray (Gy) (control), 5 Gy (single dose in the scrotal area), telmisartan, losartan, 5Gy+telmisartan and 5Gy+losartan. The treatment started the day after irradiation with losartan 34 mg/kg (two times/day) and telmisartan 12 mg/kg (one time/day) during 60 days. For ultrastructural analysis, the testis fragments were fixed in 2 % glutaraldehyde and 4 % paraformaldehyde in 0.1 M phosphate buffer, pH 7.3. The material was postfixed for 2 h in 1 % osmium tetroxide. For collagen evaluation, the sections were stained with Picrosirius-red method. Serum testosterone was determined. The date showed the deleterious effects of gamma radiation on testicular ultrastructure. Rich accumulation of collagen fibers in the interstitium was observed in the irradiated groups, especially the irradiated and nontreated testes. No significant difference was detected in serum testosterone concentration among the studied experimental groups. Treatments with telmisartan and losartan influenced the onset of attenuation on ultrastructural damages arising from ionizing radiation. Although the data strongly suggest that AT1 receptor antagonists may promote radioprotection to the testes, further studies with a longer duration of treatment are required for these potentially positive effects to be maximized and, therefore, to better characterize radioprotection to reproductive parameters.

El tratamiento radioterápico es una fuente de exposición del ser humano a la radiación ionizante. Esta energía pura causa efectos deletéreos en los tejidos, debido al estrés oxidativo, fenómeno donde hay participación del Sistema Renina-Angiotensina. Los órganos genitales masculinos son extremadamente radiosensibles y la acción de la radiación en los testículos puede afectar significativamente la espermatogénesis. En la búsqueda de potenciales radioprotectores, este estudio ha investigado fármacos antagonistas del receptor AT1 que minimizan los daños radioinduzidos en los tejidos reproductivos, por medio de la disminución del estrés oxidativo. Ratones Wistar machos fueron distribuidos en seis grupos: grupo 0 Gray (Gy) (control), grupo 5 Gy (dosis única en el área escrotal), grupo telmisartán, grupo losartán, grupo 5Gy+telmisartán y grupo 5Gy+losartán. El tratamiento empezó en el día siguiente a la irradiación con losartán 34 mg/kg (2x/día) y telmisartán 12 mg/kg (1x/día), durante 60 días. Para el análisis ultraestructural, los testículos se fijaron en glutaraldehido (2 %) y paraformaldehido (4 %) con tampón de fosfato 0,1 M, pH 7,3. El material fue post-fijado en tetróxido de osmio (1 %). Para evaluar el colágeno fue utilizado el método Picrosirius Red. Fue determinada la concentración sérica de testosterona. Los datos mostraron los efectos deletéreos de los rayos gamma sobre la ultraestructura testicular. Fue observada una rica deposición de colágeno en el intersticio en los grupos irradiados, especialmente en el irradiado y no tratado. Entre los grupos, no se detectó ninguna diferencia significativa en la concentración sérica de testosterona. Los tratamientos con telmisartán y losartán influenciaron el comienzo de la atenuación de los cambios en la ultraestructura testicular de la radiación. A pesar de que los datos sugieren que los antagonistas del receptor AT1 pueden promover radioprotección a los testículos, estudios complementarios con una duración de tratamiento más extendida son necesarios para que los efectos potencialmente positivos sean maximizados y, por supuesto, puedan mejorar la caracterizacion de la radioprotección a los parámetros reproductivos.

Reversible effect of high altitude on rat testis morphology and spermatogenesis: histological and ultrastructural study / Efecto reversible de la altitud elevada sobre la morfología y espermatogénesis en el testículo de rata: estudios histológico y ultraestructural

The study was carried out at two different altitudes in the southern region of Saudi Arabia: Abha, 2,800 meters above sea level, the high altitude (HA) area and Jazan, 40 meters above sea level the low altitude (LA) area. Following exposure to high altitude, testes of rats revealed various types of atrophy and degeneration in the seminiferous tubules and in the interstitial tissue. There was detachment of the basal laminae of the tubules and a profound decrease in cellularity. When rats were brought back to their habitat (LA) and later examined, many tubules showed normal population of cells including spermatids and spermatozoa. Well-arranged epithelium was seen in most of the seminiferous tubules of these animals, normal interstitial space and no detachment of the basal lamina. Apparently complete recovery had been achieved ultrastructurally, in hypoxic group; some spermatogenic cells lost their normal architecture, being irregular in shape with some features of necrosis, such as shrinkage and pyknotic nuclei characterized by chromatin condensation. Significant decrease in epithelial height was noticed in these animals (P <0.05). Also, the diameter of the tubules showed slight decrease with concomitant increase in interstitial spaces.

El estudio se realizó en dos ciudades con alturas diferentes en la región sur de Arabia Saudita: Abha, a 2.800 metros sobre el nivel del mar, una zona de gran altura (GA) y Jazan, a 40 metros sobre el nivel del mar, área de baja altitud (BA). Después de la exposición a una gran altura, los testículos de ratas revelaron varios tipos de atrofia y degeneración en los túbulos seminíferos y en el tejido intersticial. Hubo desprendimiento de la lámina basal de los túbulos y una disminución profunda en la celularidad. Cuando las ratas fueron devueltas a su hábitat (BA) y posteriormente examinadas, muchos túbulos mostraron un número normal de células, incluyendo espermáticas y espermatozoides. El epitelio se observó normal en la mayoría de los túbulos seminíferos de estos animales, con el espacio intersticial normal y sin desprendimiento de la lámina basal. Al parecer, se logró una recuperación ultraestructural completa en el grupo de hipoxia; algunas células de espermatogénesis perdieron su arquitectura normal, siendo de forma irregular con algunas características de necrosis, como la contracción y núcleos picnóticos caracterizados por condensación de la cromatina. Se observó disminución significativa en la altura del epitelio en estos animales (P <0,05). Además, el diámetro de los túbulos mostró una ligera disminución con aumento concomitante en los espacios intersticiales.

Histology and ultrastructure of male reproductive system of the Indian Spiny lobster Panulirus homarus (Decapoda: Palinuridae) / Histología y ultraestructura del sistema reproductor masculino de la langosta espinosa de la India Panulirus homarus (Decapoda: Palinuridae)

The spiny lobster Panulirus homarus, distributed along the Southeast and Southwest coasts of India, is an important commercial species having mariculture potential. Despite its importance, the structural and ultrastructure features of male gonads from this species have received scarce attention. Hence this study was aimed to describe the male reproductive tract of the species, using standard histological and electron microscopy techniques. Gonads from 94 specimens of P. homarus ranging in carapace length 37mm-92mm from vizhinjam (Southwest coast of India.) were obtained and processed for the study (Histology-70 numbers & ultrastructure-24 numbers). The male reproductive system consists of paired testis and vas deferens located in the cephalothoracic region. Macroscopically, the reproductive tract was observed in lobsters >35mm carapace length. In immature testis, spermatogonia were seen which measured 6.9-13.8µm in diameter and in the mature testis primary (5.4-5.9µm) and secondary spermatocytes (2.8-3µm) and spermatids (2.2-2.4µm) were present. Each vas deferens consists of proximal and distal portions. The spermatophoric mass begins formation in the proximal vas deferens. In the distal vas deferens the spermatophoric mass containing the spermatozoa are arranged in packets towards the periphery by the gelatinous matrix produced by the typhlosole. Ultrastructurally, the spermatogonia have lamina, nucleus and mitochondria like bodies, the primary spermatocytes have nucleus, dense chromatin and vacuolated cytoplasm and the spermatids have mitochondria, endoplasmic reticulum and centrioles. The endoplasmic reticulum and the nuclear envelope in the spermatids form the acrosome. The radial arms with microtubules are formed in association with the dense endoplasmic reticulum, near the nucleus. The sperm has a spherical structure with the nucleus, lamellar region, spikes and acrosome. This is the first comprehensive report of the structure of the male gametes and spermatogenesis in P. homarus from Indian waters.Rev. Biol. Trop. 62 (2): 533-541. Epub 2014 June 01.

La langosta espinosa Panulirus homarus, distribuida a lo largo de las costas sudeste y sudoeste de la India, es una especie de importancia comercial con gran potencial para la mari-cultura. A pesar de su importancia, las características estructurales y ultraestructurales de las gónadas masculinas de esta especie han sido poco estudiadas. Debido a esto, el objetivo de este estudio fue describir el aparato reproductor masculino de dicha especie, utilizando técnicas convencionales de microscopía histológica y electrónica. Se procesaron 94 ejemplares de P. homarus de vizhinjam (costa suroeste de la India) (70 individuos para histología y 24 para ultraestructura), cuyos caparazones variaron de 37 mm a 92 mm de longitud. El sistema reproductor masculino de esta especie consistió en un par de testículos y un conducto deferente situados en la región céfalo-torácica. Macroscópicamente, el aparato reproductor se observó en langostas con una longitud de caparazón >35mm. En testículos inmaduros, la espermatogonia midió 6.9-13.8μm de diámetro y se encontró presente en los testículos maduros primarios (5.4-5.9μm), espermatocitos secundarios (2.8 a 3 μm) y espermátidas (2.2-2.4μm). Cada conducto deferente consistió de porciones proximales y distales. La formación de la masa espermatofórica comienza en los conductos deferentes proximales. En el conducto deferente distal espermatofórico, la masa que contiene los espermatozoides está dispuesta en paquetes hacia la periferia, en una matriz gelatinosa producida por el tiflosol. Ultraestructuralmente, las espermatogonias presentan una lámina, núcleo y mitocondrias, los espermatocitos primarios tienen núcleo, cromatina densa y citoplasma vacuolado, mientras que las espermátidas tienen mitocondrias, retículo endoplasmático y centríolos. En las espermátidas, el retículo endoplásmico y la envoltura nuclear forman el acrosoma. Los brazos radiales con microtúbulos se forman en asociación con el retículo endoplásmico denso, cerca del núcleo. El esperma presenta una estructura esférica con el núcleo, la región laminar, las espinas y el acrosoma. Este documento constituye el primer informe exhaustivo de la estructura de los gametos masculinos y espermatogénesis en P. homarus de la India.

Possible protective effect of Moringa oleifera leaf extract on dexamethasone induced histological changes in adlt rat testes

Dexamethasone [Dex] is a widely used therapeutic agent for its immunosuppressive and anti-inflammatory action. It has adverse effects on many body organs and systems. Moringa oleifera is an antioxidant-rich natural plant. It contains vitamins C and A, and various phenolic compounds. The present study was designed to evaluate the ability of M. oleifera leaf extract to protect rat testis against Dex-induced spermatogenic defects. Thirty adult male rats were divided equally into three groups [10 animals each]: the control group [group I] and two experimental groups [groups II and III]. Rats of group II were subjected to intraperitoneal injection of 7mg/kg/day of Dex for 10 days. In group III the rats were treated with M. oleifera leaf extract at 400mg/kg/day, and then after 2h they were administered an intraperitoneal injection of Dex with the same dose as for group II for 10 days. The testes were dissected out and processed for light and electron microscope examination. Microscopic examination revealed that most of the seminiferous tubules of group II were lined with germ cells with dark pyknotic nuclei and vacuolated cytoplasm. The lumen of some tubules was obliterated with exfoliated and sometimes multinucleated giant cells. There was statistically highly significant increase in the percentage of sperm abnormality. Degenerated interstitial Leydig cells were also observed. However, in the moringa-treated group, the histological changes were reduced and the percentage of sperm abnormality was more or less similar to that of the control group. These results demonstrated that M. oleifera leaf extract has a potent protective effect against the testicular toxicity induced by Dex and hence might be clinically useful

Effect of cyclosporine-A on the structure of adult albino rat testis and the role of lycopene supplementation: a histological and immunohistochemical study

Cyclosporine A [CsA] has improved the quality of life and survival rate of transplant patients. However, the oxidative stress induced by CsA limits its use as it causes side effects in different organs. Lycopene is an antioxidant found in tomatoes, water melon, and pink guava. This study aimed to investigate the histological and immunohistochemical changes occurring in rat testes following CsA intake and evaluate the role of lycopene supplementation. Thirty adult male albino rats were divided into three groups: group I, group II, and group III. Group I was divided into subgroup IA, which received distilled water, and subgroup IB, which received lycopene [10 mg/kg/day] dissolved in corn oil. Group II received CsA [15 mg/kg/day] dissolved in distilled water and group III received CsA+lycopene at the same doses as above. All treatments were given by oral gavage for 21 successive days. Testis samples were prepared for light microscopic [histological and immunohistochemical] and electron microscopic examination. The area percentage of bcl-2 reaction and height of the germinal epithelium were morphometrically measured and statistically analyzed. The germinal epithelial cells of the CsA-treated group were separated and were seen to contain dark nuclei. Sloughed germ cells in the lumen were seen. Ultrastructurally, primary spermatocytes showed vacuoles. Spermatids had shrunken nuclei and irregular distribution of mitochondria. Leydig cells contained lipid droplets of different densities with projecting multiple processes. The basement membrane was thick and contained multiple collagen fibers. The axonemes of the mid-pieces of sperms were disorganized with swollen mitochondrial sheathes. Statistically, the area percentage of bcl-2 reaction and germinal epithelial height showed a significant decrease in group II versus other groups. Lycopene improved the adverse effects of CsA in group III. CsA induced profound damage in the testicular structure in rats. It was ameliorated by concomitant lycopene administration. Thus, these results could be considered for further clinical investigations to recommend lycopene with CsA in transplant patients

Effect of zinc-free diet from weaning through puberty on the structure of the testis in albino rats, with a special focus on the Leydig cells

Zinc is an essential trace element. Many physiological processes would be impaired if zinc is not supplied in sufficient quantities in the diet. The aim of the work was to evaluate the effect of a zinc-free diet from weaning through puberty on the testicular structure of rats, with a special focus on the Leydig cells. Twenty weaned rats were divided into three groups. Group I [the control group] was fed a balanced diet. Group II [the zinc-free group] was fed a zinc-free diet for 3 months. Group III [the recovery group] was fed a zinc-free diet for 3 months, followed by a balanced diet for 1 month. At the end of the experiment, rats were weighed and blood samples were collected to measure the level of serum testosterone hormone. The testes were removed, weighed, and processed for light and transmission electron microscopic study. The zinc-free group showed a significant decrease in the mean body weight, testis weight, and serum testosterone level compared with the control group. The light and electron microscopic examination showed loss of most of the spermatogenic cells accompanied by variable degrees of degeneration in the form of karryorhexis and haphazard chromatin content. In the interstitial tissue, there was accumulation of exudate. Many Leydig cells showed dilated smooth endoplasmic reticulum and mitochondria. Others cells showed accumulation of glycogen. Most of these changes were reversed after receiving the balanced diet in the recovery group. A zinc-free diet altered the structure of both the seminiferous tubules and the Leydig cells. Most of these alterations were reversed by zinc replenishment. Zinc is considered an essential element for maintaining testicular structure and spermatogenesis process

Protective effect of orange and grapefruit peel extracts against testicular toxicity induced by sulfasalazine in male albino rats

Sulfasalazine is a drug commonly used for the treatment of inflammatory bowel diseases such as ulcerative colitis and Crohn's disease. However, the histological changes in the testes are not well defined. Also, orange and grapefruit peels are powerful antioxidants that have come into use recently for the treatment of infertility. This study aimed to elucidate the histological, immunohistochemical, and ultrastructural changes in the testes after sulfasalazine treatment and evaluate the possible protective role of orange and grapefruit peel extracts. This study included 48 adult male albino rats divided into six equal groups: the control group [group I]; the orange peel extract group [group II]; the grapefruit peel extract group [group III]; the sulfasalazine group [group IV]; the sulfasalazine and orange peel extract group [group V]; and the sulfasalazine and grapefruit peel extract group [group VI]. At the end of the experiment [2 weeks], all animals were sacrificed and their testes were excised. Paraffin sections were prepared and stained with H and E and immunohistochemical staining was performed for proliferating cell nuclear antigen. Other pieces of the testis were used for ultrastructural study. Sulfasalazine was shown to affect the testes. The changes were in the form of irregular degenerated seminiferous tubules, germ cells, decrease in proliferating cell nuclear antigen, ultrastructural alterations of spermatogenic cells, and thickened basement membranes. These changes were present in some tubules in the testes. Treatment with orange or grapefruit peel extracts proved to improve these changes. Sulfasalazine has deleterious effects on the structure of the testes and supplementation with orange or grapefruit peel extracts with sulfasalazine can overcome the toxicity of sulfasalazine on the testis and protect testicular tissue from the detrimental effects of sulfasalazine

Assessment of the protective role of vitamine E in atrazine toxicity on the testis of adult albino rats: a histological and biochemical study

Atrazine [ATZ] is one of the most commonly used herbicides that adversely affect the reproductive system in rats. The aim of the study was to evaluate the impact of vitamin E [Vit E] on subchronic exposure to ATZ in testicular tissue. Fifty adult male albino rats were divided into five groups: the negative control group; the positive control group, which received 1 ml of corn oil/day, orally; the Vit E group, which received Vit E at 100 mg/kg/day, orally; the ATZ group, which received ATZ at 300 mg/kg/day, orally; and the ATZ+Vit E group, which received both ATZ and Vit E at the previously mentioned doses. Treatments were given for 6 days/week for 45 days. At the end of the experiment, blood samples were taken to measure serum testosterone level. Semen analysis and estimation of oxidative stress markers catalase, superoxide dismutase, and tissue malondialdehyde were carried out. The testes were processed for light and electron microscopic examination. An immunohistochemical study was performed for detection of Bcl2. ATZ caused a decrease in serum testosterone level and in superoxide dismutase and catalase enzyme activities, whereas malondialdehyde content increased. There was also a decrease in sperm count, viability, and motility in comparison with the control groups. Light microscopic examination of seminiferous tubules revealed degeneration of the germinal epithelium. The lumen contained sloughed cells and homogenous acidophilic material. Ultrastructurally, there was separation of the germinal epithelial cells with small dense nuclei and phagocytic remnants. Sertoli and Leydig cells were also affected. Bcl-2 immunolocalization revealed weak reaction in the cytoplasm of the germinal epithelial cells and Leydig cells. Concomitant use of Vit E caused partial improvement. ATZ induced detrimental effects in the testicular tissue that were attenuated by concomitant administration of vitamin E

Role of ginseng on mercury chloride-induced testicular lesions in adult albino rat: a histological and immunohistochemical study

Mercury is a toxic metal causing various adverse neurological, developmental, and reproductive disorders. Mercury exposure occurs through contaminated water and food. Ginseng [herbal medicine] can improve male fertility by acting directly on the pituitary gland and central nervous system. The aim of the study was to evaluate the role of Ginseng on mercury chloride-induced testicular toxicity in adult albino rats. Thirty-two adult male albino rats were divided randomly into four groups of eight rats each: group I [the Control group]: the animals in this group received 1.5 ml of distilled water; group II [the Ginseng group]: the animals in this group received 100 mg/kg body weight of Ginseng; group III [the Mercury Chloride group]: the animals in this group received mercury chloride at a dose of 2.0 mg/kg/day. Group IV [the Ginseng and Mercury Chloride group]: the animals in this group received both Ginseng and mercury chloride at the same dose and route of administration as the previous group. The testes were removed and fixed. Paraffin sections were prepared and stained with H and E and Masson's trichrome, and immunohistochemical staining was performed for proliferating cell nuclear antigen. Morphometric study was conducted and results were statistically analyzed. The study showed significant increase in body weight in group II and decrease in body weight in groups III and IV compared with the Control group. The gonadal index of group III showed significant decrease compared with both the Control group and group II. H and E sections of group III showed loosely packed vacuolated seminiferous tubules. Some germ cells showed a hyperchromatic nucleus, whereas others showed chromatolysis. Absence of mature sperms was noticed. Some tubules showed massive loss of germ cells. In the treated group [group IV] germ cells regained more or less a normal appearance. Ginseng can ameliorate the toxic changes of mercury chloride on the rat testis

Histological and biochemical evaluation of the effects of some antioxidants on aged testes

Different studies have demonstrated that aging clearly affects male fertility which may be attributed to the androgen deficiency. Reactive oxygen species play a central role in the pathophysiology in the aged-related decrease in male fertility. Some antioxidants have ameliorative effects on different aged organs. The present study aimed to evaluate the effects of some antioxidants on aged testes. Ten adult and fifty aged male albino rats [Rattus albus] were divided into six groups. Group I [control adult], Group II [control aged], Group III [Vitamin E-treated aged], Group IV [Vitamin C-treated aged], Group V [Zinc sulphate-treated aged], Group VI [Vitamin E-, Vitamin C-and Zinc Sulphate-treated aged]. Vitamin E, Vitamin C and Zinc were administrated in doses 2.52 mg, 3.15 mg and 0.693 mg, respectively. Histological and ultrastructural evaluation of the testes were examined as well as Follicle stimulating hormone [FSH], Luteinizing hormone [LH], total and free testosterone levels in the serum were measured. Counting the number of litters per animal and the teratogenic effects was noticed. Giving zinc alone or combined with other antioxidants gave better ameliorative effects on the testicular structure and hormonal levels in the serum. No teratogenic effects of the aged animals`offspring were noticed

Long term evaluation of morphometric and ultrastructural changes of testes of alloxan-induced diabetic rats

PURPOSE: To evaluate in a long term the morphometric and ultrastructural changes in seminiferous tubules (ST) of normal and diabetic rats, and to correlate any changes with animal age and diabetes duration. METHODS: Sixty male Wistar rats, three months-old, were randomly divided into two groups: 30 non-diabetic controls (N) and 30 alloxan untreated diabetic (D). After one, six and 12 months of follow-up or diabetes induction rats were sacrificed and the testes examined. Morphometric measures of the ST were performed by digital imaging analysis. ST ultrastructure was analyzed by transmission electron microscopy. RESULTS: Sustained hyperglycemic state was observed in all diabetic rats throughout the study. Serum testosterone was also significantly decreased in these animals. The diameter, total area, epithelium area and epithelium thickness of ST were lower and tubular density was higher in diabetic animals. Diabetic rats also showed ultrastructural changes compromising the whole testis including germ-, Sertoli-, and Leydig cells, and also the mithocondria and cellular nuclei. Most frequent of these consisted of vacuolization and/or accumulation of lipid droplets and electron dense dark material in cell cytoplasm and/or in membranes, cellular degeneration, and apoptosis. Non-diabetic control rats also showed testicular lesions that resemble to the diabetic lesions, although much less severe and with later onset in life of these animals. CONCLUSION: Histopathological changes observed in testes of normal and diabetic rats are closely related to the animal age and/or duration of the hyperglycemic state, being progressively more severe in animals sacrificed belatedly. These changes may play an important role in male infertility observed in diabetic subjects.

possible correlation between the testicular structure and short photoperiod exposure in young albino rats: light and electron microscopic study

Light is considered as the first ecological factor affecting primary productivity. Several studies have attempted to relate between seasonal changed environmental light factors and the reproductive system. The influence of environmental light on the anatomy and physiology of different organs has been investigated intensely in a different species. Therefore, this research was designed to examine the effects of a short duration of light exposure on the histological structures of rat testes. Eighteen young male albino rats were divided into control [I] and experimental [II] groups. They were kept for 8 weeks in separate cages. The rats in group I were exposed daily to a normal lighting cycle of 12 h light and 12 h darkness. Light was provided by a 20-W 4-ft fluorescent lamp. Group II was exposed daily to 4 h of light and 20 h of darkness. They were housed in a small room. Its windows were covered with a black cotton material. At the end of the experiment, all rats were anaesthetized with ether, and their testes were dissected out and processed for light and electron microscopic examination. In group II, spermatogenic cells of some seminiferous tubules appeared separated and exfoliated. The other tubules were destroyed, with acidophilic hyaline material. Loss of germ cells was confirmed by a significantly low sperm count. Some germ cells showed a vacuolated cytoplasm and disrupted intercellular bridges with the formation of giant cells. Sertoli cells showed active phagocytic capacity with the appearance of multivesicular bodies. Hormonal assay showed a low significant testosterone level; this was supported by the presence of inactive spindle-shaped Leydig cells. The present study confirmed that rats were structurally and functionally photosensitive. Therefore, the changes in the normal photoperiod could influence their reproductive functions

Role of coenzyme Q10 in testicular damage induced by acrylamide in weaned albino rats: a histological and immunohistochemical study

Acrylamide is a vinyl monomer frequently used in the polymer industry. It has the potential to adversely affect male reproductive capacity. Coenzyme Q10 [CoQ10] is a strong antioxidant. The objectives of this study were to examine the histological changes in the testis after the administration of acrylamide and the possible protective role of CoQ10. Thirty weaned male albino rats aged 21 days were classified into three groups of 10 rats each: group I was the control group; group II [acrylamide-treated rats] received oral acrylamide at 15 mg/kg body weight/day; and group III [protected group] received both acrylamide [at the same previous dose] and an intrsperitoneal injection of CoQ10 at 10 mg/kg body weight/day. After 8 weeks, blood samples were taken to measure the serum testosterone level. The testes from each animal were dissected out and processed for light microscope examination using Hand E stains and immunohistochemical stains for the detection of proliferating cell nuclear antigen and inducible nitric oxide synthase. Morphometric and statistical analyses were carried out. After the administration of acrylamide, variable degrees of tubular affection were observed. Some tubules were shrunken with disorganized germinal epithelium. Spermatogonia contained darkly stained nuclei. Cellular vacuolations as well as sloughed spermatogenic cells into the lumen were observed. The interstitium was widened with interstitial hyperplasia, eosinophilic material, and congested capillaries. Immunohistochemically, acrylamide treatment induced a marked reduction in the number of proliferating cell nuclear antigen immunoreactive Spermatogonia and spermatocytes and an increase in the number of inducible nitric oxide synthase immunoreactive spermatids and spermatocytes. The concomitant administration of CoQ10 with acrylamide induced an observable protection against these changes. CoQ10 played a protective role against acrylamide-induced testicular damage

Testicular toxic changes induced by bisphenol A in adult albino rats: a histological, biochemical, and immunohistochemical study

Bisphenol A [BPA] is an endocrine disruptor that is incorporated in many plastic industries worldwide. The exposure of humans to such substances starts from the fetal life to the postnatal life and extends throughout the life of the individual. Many agencies have raised warnings against the excessive use of such substances. The present study was designed to evaluate the biochemical and histological changes induced by BPA in the testis of adult male albino rats and to detect the ability of self-regeneration after stoppage. Thirty-two adult male albino rats were used. The rats were divided equally into four groups [eight animals each]. Groups I and II were used as negative and positive control groups, respectively. Rats of group III were given an oral dose of 50 mg/kg of BPA per day for 8 weeks. In group IV, the rats were treated in the same manner as in group III and then left without treatment for 4 weeks for recovery. At the time of sacrifice, all rats were anesthetized with ether, and blood samples were collected for estimation of testosterone. The testes were dissected out and processed for testicular malondialdehyde and glutathione measurement and light and electron microscopic examination. The diameter and epithelial height of the seminiferous tubules were estimated morphometrically and statistically analyzed. Biochemical results of the BPA-treated group [group III] revealed testicular affection with oxidative stress. Testes of this group showed many distorted seminiferous tubules lined by disorganized epithelium and separated with wide interstitium containing congested blood vessels. Apoptotic nuclei of some spermatids and intercellular spaces were also seen. There was a decrease in estrogen receptors. Statistical analysis of epithelial height and tubular diameter confirmed the results. However, in the recovery group [group IV], the histological and the biochemical changes were reduced but did not return to normal. These results demonstrated that BPA had deleterious effects on the testis with some sort of self-recovery after stoppage

Histological study on the role of ginger against cisplatin-induced testicular toxicity in albino rats

Chemotherapy with cisplatin has adverse effects on spermatogenesis. Therefore, this work aimed at investigating the protective role of ginger against cisplatin-induced testicular toxicity in male albino rats. Twenty-four adult albino rats were used in this study. They were divided into three groups. The first group served as the control group; the second group was injected with cisplatin [12 mg/kg once]; and the third group was injected with cisplatin [12 mg/kg once] and then given ginger [310 mg/kg orally] for 26 days. Testicular specimens were processed for light microscopic examination using H and E. Other specimens were processed for electron microscopic examination. Cisplatin had damaging effects on the seminiferous tubules. Some areas of the tubules showed complete depletion of germ cells. Other areas showed some spermatogonia or primary spermatocytes. Sertoli cells showed a variable degree of degenerative changes in the form of destruction of cellular processes and cell junction. Interruption of the nuclear envelope of spermatids and loss of intercellular bridges were noticed. Treating with ginger resulted in normal Sertoli cells and cell junctions. The germ cells lining the tubules were more or less normal except for some intercellular vacuolations. The use of ginger has some protective effects on the testicular structure; hence, a larger number of experiments with higher doses of ginger or longer administration period could be beneficial for patients taking chemotherapeutic drugs

adverse effects of bisphenol 'A' on some reproductive organs of the male albino rat: a light and electron microscopic study

Background: Bisphenol A [BPA] is a xenoestrogen [environmental estrogens] used in the production of polycarbonate plastics and epoxy resins that line food and beverage cans. Modernization of the Arabian Gulf region has resulted in the wide consumption of readymade foods that are packed in plastic containers and cans. Consequently, the majority of humans, particularly infants and children, are being continuously exposed to it

Aim of work: The objective of the present study was to examine the effect of exposure to BPA on the testis, epididymis, prostate, and penile corpora of adult albino rat

Materials and methods: One, 4, and 8 weeks following a subcutaneous injection of 150 microg BPA/kg body weight into adult albino rats every other day for 12 days, the histopathological changes induced in the testis, epididymis, prostate, and penile corpora were detected using both light and transmission electron microscopic techniques

Results: In BPA-treated animals, seminiferous tubules showed a decreased thickness of germinal epithelium with vacuolar degeneration and increased apoptotic cells. Sperm were hardly seen till the eighth week, when spermatogenesis was regained, but spermatids and mature sperm still had residual malformations. The rough endoplasmic reticulum cisternae of prostatic parenchyma appeared distended with a homogeneous content whereas most of the secretory vesicles were empty. In the penile corpora of BPA-treated groups, both tunical thickness and trabecular smooth muscle content were increased with consequent narrowing of sinusoidal spaces

Conclusion: These results suggest that BPA inhibits spermatogenesis, increases the ratio of sperm anomalies, and has a potential harmful effect on erectile function, which raises an alarm to the harmful effects of environmental contaminants that might cause subfertility or infertility

Ultrastructure of Spermatogenesis in the Testis of Paragonimus heterotremus

Lung fluke, Paragonimus heterotremus, is a flatworm causing pulmonary paragonimiasis in cats, dogs, and humans in Southeast Asia. We examined the ultrastructure of the testis of adult P. heterotremus with special attention to spermatogenesis and spermiogenesis using scanning and transmission electron microscopy. The full sequence of spermatogenesis and spermiogenesis, from the capsular basal lamina to the luminal surface, was demonstrated. The sequence comprises spermatogonia, spermatocytes with obvious nuclear synaptonemal complexes, spermatids, and eventual spermatozoa. Moreover, full steps of spermatid differentiation were shown which consisted of 1) early stage, 2) differentiation stage representing the flagella, intercentriolar body, basal body, striated rootlets, and electron dense nucleus of thread-like lamellar configuration, and 3) growing spermatid flagella. Detailed ultrastructure of 2 different types of spermatozoa was also shown in this study.