The study examined the role of endoplasmic reticulum stress (ERS) and signaling pathways of inositol-requiring enzyme-1 (IRE1), RNA-activated protein kinase-like ER kinase (PERK) and activating transcription factor-6 (ATF6) in apoptosis of mouse testicular cells treated with low-dose radiation (LDR). In the dose-dependent experiment, the mice were treated with whole-body X-ray irradiation at different doses (25, 50, 75, 100 or 200 mGy) and sacrificed 12 h later. In the time-dependent experiment, the mice were exposed to 75 mGy X-ray irradiation and killed at different time points (3, 6, 12, 18 or 24 h). Testicular cells were harvested for experiments. H(2)O(2) and NO concentrations, and Ca(2+)-ATPase activity were detected by biochemical assays, the calcium ion concentration ([Ca(2+)]i) by flow cytometry using fluo-3 probe, and GRP78 mRNA and protein expressions by quantitative real-time RT-PCR (qRT-PCR) and Western blotting, respectively. The mRNA expressions of S-XBP1, JNK, caspase-12 and CHOP were measured by qRT-PCR, and the protein expressions of IRE1α, S-XBP1, p-PERK, p-eIF2α, ATF6 p50, p-JNK, pro-caspase-12, cleaved caspase-12 and CHOP by Western blotting. The results showed that the concentrations of H2O2 and NO, the mRNA expressions of GRP78, S-XBP1, JNK, caspase-12 and CHOP, and the protein expressions of GRP78, S-XBP1, IRE1α, p-PERK, p-eIF2α, ATF6 p50, p-JNK, pro-caspase-12, cleaved caspase-12 and CHOP were significantly increased in a time- and dose-dependent manner after LDR. But the [Ca(2+)]i and Ca(2+)-ATPase activities were significantly decreased in a time- and dose-dependent manner. It was concluded that the ERS, regulated by IRE1, PERK and ATF6 pathways, is involved in the apoptosis of testicular cells in LDR mice, which is associated with ERS-apoptotic signaling molecules of JNK, caspase-12 and CHOP.
Animals ; Apoptosis ; physiology ; radiation effects ; Endoplasmic Reticulum Stress ; physiology ; radiation effects ; Male ; Mice ; Radiation ; Testis ; physiology ; radiation effects

The testis is highly sensitive to electromagnetic radiation. Sperm is the passer of male genetic material and electromagnetic radiation may cause structural and functional injury to the testis, including motility reduction, abnormality increase and ultrastructural alteration of epididymal sperm. Energy metabolism disorder in spermatogenic cells, enhancement of lipid peroxidation in the testis, excessive expression of inflammatory factors and abnormality of genetic transcription may be responsible for injury to the testis and epididymal sperm. This paper reviews the progress made in this field and the preventive measures against the injury.
Animals ; Epididymis ; radiation effects ; Humans ; Male ; Radiation ; Rats ; Rats, Sprague-Dawley ; Sperm Count ; Sperm Motility ; radiation effects ; Testis ; radiation effects

OBJECTIVETo study the effect of high power microwave (HPM) radiation on the testicular germ cell apoptosis.

METHODSOne hundred and twenty-five Spraque-Dawley rats were randomly divided into two groups, unexposed control group and experimental group(further divided into four subgroups: 10 mW/cm2 5 min, 10 mW/cm2 10 min, 20 mw/cm2 5 min, and 20 mW/cm2 10 min), and then the experimental group was radiated with S wave band of 10 mW/cm2, 20 mW/cm2 high power microwave for 5 or 10 min. Testicular samples were taken at 6 h, 24 h, 48 h, 72 h and 5 d after radiation and separately studied. At the end of the process, testicular germ cell apoptosis was detected by in situ terminal deoxynucleotityl transferase mediated dUTP nick end labeling (TUNEL).

RESULTSThe number of apoptotic cells of the 6 h, 24 h and 48 h experimental groups at 5 min after 10 and 20 mW/cm2 radiation was remarkably larger than that of the controls (P < 0.01), especially after 10 mW/cm2 radiation, the number of the 6 h group reached the peak (161.27 +/- 5.90) /5 convoluted seminiferous tubules. The changes in the other experimental groups had no significant difference compared with the controls (P > 0.05).

CONCLUSIONHPM can increase germ cell apoptosis of the rat testis, which is related to the time of radiation and sample acquisition. In the condition of the present test, 5 minutes of HPM radiation may significantly enhance testicular germ cell apoptosis and damage, which in turn may influence the reproductive function of the rats.

Animals ; Apoptosis ; radiation effects ; Male ; Microwaves ; Rats ; Rats, Sprague-Dawley ; Spermatozoa ; pathology ; radiation effects ; Testis ; radiation effects

OBJECTIVETo explore the pathological characteristics and the dynamic change regularity of the testis induced by high power microwave (HPM) radiation.

METHODSOne hundred and sixty-five male Wistar rats were exposed to 0, 3, 10, 30 and 100 mW/cm2 HPM radiation for five minutes, and changes of testicular morphology and teratogenic ratio of epididymal spermatozoa were observed through light microscope and electron microscope at 6 h, 1, 3, 7, 14, 28 and 90 d after radiation.

RESULTSInjury of testicular spermatogenic cells in rats might be induced by 3 to approximately 100 mW/cm2 HPM radiation, and the main pathological changes were degeneration, necrosis, shedding of spermatogenic cells, formation of multinuclear giant cells, decrease or loss of sperm and interstitial edema. Injury of spermatogenic cells underwent such phases as death and shedding, cavitation, regeneration and repair, characterized by being focalized, inhomogenous and phased. And the severity of pathological changes of the testis increased with power density. There was only scattered degeneration, necrosis, shedding of spermatogenic cells in the seminiferous tubule one day after 3 mW/cm2 radiation, and the pathological changes six hours after 10 mW/cm2 radiation was similar to those one day after 3 mW/cm2 radiation, but with the formation of multinuclear giant cells, and the above-mentioned pathological changes aggravated from one day to seven days after radiation. There was a significant increase in degeneration, necrosis, shedding of spermatogenic cells, as well as a significant decrease in spermatozoa and focal necrosis in simple seminiferous tubules six hours after 30 and 100 mW/cm2 radiation, and the subsequent changes were similar to those of 10 mW/cm2 radiation. There was a significant increase in teratogenic ratio of epididymal spermatozoa at 3 d, 1 to approximately 7 d, 6 h to approximately 7 d after 3, 10, 30 and 100 mW/cm2 microwave radiation respectively (P < 0.01 or P < 0.05).

CONCLUSIONHPM radiation may cause injury of testicular spermatogenic cells in rats, which has a positive correlation to radiation dosage and time.

Animals ; Dose-Response Relationship, Radiation ; Male ; Microwaves ; Rats ; Rats, Wistar ; Spermatozoa ; pathology ; radiation effects ; Testis ; pathology ; radiation effects

ObjectiveTo investigate the influence of cellphone electromagnetic radiation (CER) on the testicular ultrastructure and the apoptosis of spermatogenic cells in male rats.atability, feasibility, applicability, and controllability in the construction of experimental animal models, we compared the major anatomic features of the penis of 20 adult beagle dogs with those of 10 adult men. Using microsurgical techniques, we performed cross-transplantation of the penis in the 20 (10 pairs) beagle dogs and observed the survival rate of the transplanted penises by FK506+MMF+MP immune induction. We compared the relevant indexes with those of the 10 cases of microsurgical replantation of the amputated penis.

METHODSThirty adult male SD rats were equally randomized into a 2 h CER, a 4 h CER, and a normal control group, the former two groups exposed to 30 days of 900 MHz CER for 2 and 4 hours a day, respectively, while the latter left untreated. Then the changes in the ultrastructure of the testis tissue were observed under the transmission electron microscope and the apoptosis of the spermatogenic cells was determined by TUNEL.

RESULTSCompared with the normal controls, the rats of the 2 h CER group showed swollen basement membrane of seminiferous tubules, separated tight junction of Sertoli cells, increased cell intervals, apparent vacuoles and medullization in some mitochondria, and increased apoptosis of spermatogenic cells, mainly the apoptosis of primary spermatocytes (P<0.05 ). In comparison with the 2 h CER group, the animals of the 4 h CER group exhibited swollen basement membrane of seminiferous tubules, more separated tight junction of Sertoli cells, wider cell intervals, incomplete membrane of spermatogonial cells, fragments of cytoplasm, nuclear pyknosis and notch, slight dilation of perinuclear space, abnormalities of intracellular mitochondria with vacuoles, fuzzy structure, and fusion or disappearance of some cristae, and increased damage of mitochondria and apoptosis of spermatogenic cells, including the apoptosis of spermatogonial cells, primary spermatocytes, and secondary spermatocytes (P<0.05 ).

CONCLUSIONSCER can damage the testicular ultrastructure and increase the apoptosis of spermatogenic cells of the male rat in a time-dependent manner, and the apoptosis of spermatogenic cells may be associated with the damage to mitochondria.

Animals ; Apoptosis ; Cell Phone ; Electromagnetic Radiation ; Male ; Mitochondria ; radiation effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Seminiferous Tubules ; radiation effects ; Sertoli Cells ; radiation effects ; Spermatocytes ; radiation effects ; Spermatogonia ; radiation effects ; Testis ; radiation effects ; ultrastructure

OBJECTIVETo investigate the effects of extremely low frequency electromagnetic fields (ELF EMFs) on male reproduction in mice.

METHODS94 adult male mice were exposed to 50 Hz sinusoidal electromagnetic fields of 0.2, 3.2 or 6.4 mT for 2 weeks or 4 weeks. Testicular histology and weight, sperm amount, sperm motility and morphology were measured. The percentages of different ploidy cells and cell phases, and DNA content of testis cells were estimated by flow cytometry. The micronucleus rate of bone-marrow cell was also observed.

RESULTSThe testicular weight of the mice exposed to 6.4 mT for 4 weeks [(76.06 +/- 32.25) mg] was significantly lower than that of the control [(111.44 +/- 19.99) mg, P < 0.05]; no significant histopathological changes were observed on the testis in EMFs exposed mice;the sperm amount was decreased after EMFs exposure for 4 weeks, and those of the mice exposed to 0.2 mT and 6.4 mT for 4 weeks [(4.87 +/- 0.94) x 10(6)/ml and (4.30 +/- 1.89) x 10(6)/ml respectively] were significantly lower than that of the control [(6.67 +/- 0.70) x 10(6)/ml, P < 0.05]; the rates of sperm motility also showed a decline. After 0.2, 3.2 or 6.4 mT EMFs exposure for 2 weeks, the deformity rates of sperm [(7.416 +/- 3.352)%, (6.862 +/- 2.947)% and (8.112 +/- 4.615)% respectively] were significantly higher than that of the control [(4.098 +/- 2.028)%, P < 0.01]. Similarly, those of the mice exposed for 4 weeks [(10.267 +/- 3.836)%, (11.027 +/- 7.059)%, (8.814 +/- 3.678)% respectively] were higher than that of the control [(3.714 +/- 1.830)%]. After 6.4 mT exposure for 2 weeks, the percentages of 1C testis cells [(69.56 +/- 4.07)%] was significantly lower than that of the control [(73.45 +/- 3.10)%, P < 0.05]. There were not any remarkable changes in those of 2C, 4C cells. DNA content in different ploidy cells of the mice exposed to 6.4 mT was decreased. Moreover, the cell percentage in S phase was increased significantly (P < 0.01).

CONCLUSIONELF EMFs exposure may have some adverse effects on reproduction in mice.

Animals ; DNA ; metabolism ; Electromagnetic Fields ; Male ; Mice ; Random Allocation ; Reproduction ; radiation effects ; Sperm Count ; Sperm Motility ; radiation effects ; Spermatozoa ; cytology ; metabolism ; radiation effects ; Testis ; cytology ; radiation effects

OBJECTIVETo investigate the effect of long-term microwave radiation on male reproduction in rats.

METHODSA total of 100 male Wistar rats were exposed to microwave radiation with average power density of 0, 2.5, 5 and 10 mW/cm2 for 4 weeks, 5 times a week and 6 minutes per time. Changes in serum testosterone, testicular index, histology and ultrastructure, and the percentage of teratospermia in the epididymis were observed dynamically at 6 h, 7 d, 14 d, 28 d and 60 d after the exposure.

RESULTSThere was a significant decrease in serum testosterone concentration at 28 d after microwave radiation at 2.5, 5 and 10 mW/cm2 ([10.20 +/- 4.31] ng/ml, [5.56 +/- 3.47] ng/ml and [7.53 +/- 4.54] ng/ml) and at 60 d at 10 mW/cm2 ( [15.95 +/- 9.54] ng/ml), as compared with the control group ([23.35 +/- 8.06] ng/ml and [31.40 +/- 9.56] ng/ml) (P < 0.05 or P < 0.01). No significant changes were found in the testis index at 6 h -60 d after microwave radiation at the three doses, but different degrees of degeneration, necrosis and shedding of spermatogenic cells, thinning of spermatogenic epithelia, and decrease or deletion of spermatozoa were observed, and more obvious at 28 d and 60 d. Swelling and cavitation of mitochondria in all spermatogenic cells, agglutination and margin translocation of nuclear chromatin in the spermatogonial and Leydig cells were seen at 7 d and 60 d after 5 mW/cm2 microwave radiation. The rate of teratospermia of the epididymis was increased, more obviously at 7 d after 2.5, 5 mW/cm2, 60 d after 5 mW/cm2, and 7 d, 28 d and 60 d after 10 mW/cm2 microwave radiation (P < 0.05 or P < 0.01).

CONCLUSIONLong-term microwave radiation may cause injury to male reproduction, which is positively correlated with the radiation dose, and has an obvious late effect.

Animals ; Dose-Response Relationship, Radiation ; Male ; Microwaves ; adverse effects ; Rats ; Rats, Wistar ; Reproduction ; radiation effects ; Sperm Head ; radiation effects ; Testis ; radiation effects

With the popularized use cell phones, more and more concern has been aroused over the effects of their radiation on human health, particularly on male reproduction. Cell phone radiation may cause structural and functional injuries of the testis, alteration of semen parameters, reduction of epididymal sperm concentration and decline of male fertility. This article presents an overview on the impact of cell phone radiation on male reproduction.
Cell Phone ; DNA Damage ; Epididymis ; radiation effects ; Humans ; Male ; Oxidative Stress ; Semen ; radiation effects ; Sperm Count ; Sperm Motility ; radiation effects ; Testis ; radiation effects

OBJECTIVETo investigate the effect of simulated microgravity and carbon ion irradiation (CIR) on spermatogenic cell apoptosis and sperm DNA damage to the testis of male Swiss Webster mice, and assess the risk associated with space environment.

METHODSSperm DNA damage indicated by DNA fragmentation index (DFI) and high DNA stainability (HDS) was measured by sperm chromatin structure assay (SCSA). Apoptosis of spermatogenic cells was detected by annexin V-propidium iodide assay. Bax (the expression levels of p53) and proliferating cell nuclear antigen (PCNA) were measured by immunoblotting; p53 and PCNA were located by immunohistology.

RESULTSHDS, DFI, apoptosis index, and the expression levels of p53 and Bax were detected to be significantly higher in the experimental groups (P<0.05) compared with those in the control group; however, the PCNA expression varied to a certain degree. p53- and PCNA- positive expression were detected in each group, mainly in relation to the spermatogonic cells and spermatocytes.

CONCLUSIONThe findings of the present study demonstrated that simulated microgravity and CIR can induce spermatogenic cell apoptosis and sperm DNA damage. Sperm DNA damage may be one of the underlying mechanisms behind male fertility decline under space environment. These findings may provide a scientific basis for protecting astronauts and space traveler's health and safety.

Animals ; Apoptosis ; radiation effects ; Carbon ; Cell Proliferation ; radiation effects ; DNA Damage ; Heavy Ions ; adverse effects ; Immunohistochemistry ; Male ; Mice ; Random Allocation ; Sperm Count ; Spermatogenesis ; radiation effects ; Spermatozoa ; radiation effects ; Testis ; radiation effects ; Weightlessness Simulation

OBJECTIVETo determine the effect of high power microwave (HPM) radiation on the structure and function of blood-testis barrier (BTB) in rats.

METHODSOne hundred and sixty-six male Wistar rats were treated by heart perfusion of lanthanum-glutaraldehyde solution and tail vein injection of evans blue (EB) at 6 h, 1, 3, 7 and 14 d after exposed to 0, 10, 30 and 100 mW/cm2 HPM radiation for 5 minutes, the structural change of BTB and distribution of lanthanum or EB observed through the light microscope, electron microscope and laser scanning confocal microscopy (LSCM).

RESULTSTesticular interstitial edema, vascular congestion or hyperemia with accumulation of plasma proteins and red blood cells in the inner compartment of seminiferous tubules were observed after exposure to HPM. The above-mentioned pathological changes were aggravated at 1-7 d and relieved at 14 d after radiation, obviously more severe in the 30 and 100 mW/cm2 exposure groups than in the 10 mW/cm2. Both lanthanum precipitation and EB were deposited in the inner compartment.

CONCLUSIONHPM radiation may damage the structure and increase the permeability of BTB.

Animals ; Blood-Testis Barrier ; pathology ; physiopathology ; radiation effects ; Male ; Microwaves ; adverse effects ; Rats ; Rats, Wistar