simple technique for three-dimensional imaging and segmentation of brain vasculature u sing fast free-of-acrylamide clearing tissue in murine

Objective: Fast Free-of-Acrylamide Clearing Tissue [FACT] is a recently developed protocol for the whole tissue three-dimensional [3D] imaging. The FACT protocol clears lipids using sodium dodecyl sulfate [SDS] to increase the penetration of light and reflection of fluorescent signals from the depth of cleared tissue. The aim of the present study was using FACT protocol in combination with imaging of auto-fluorescency of red blood cells in vessels to image the vasculature of a translucent mouse tissues

Materials and Methods: In this experimental study, brain and other tissues of adult female mice or rats were dissected out without the perfusion. Mice brains were sliced for vasculature imaging before the clearing. Brain slices and other whole tissues of rodent were cleared by the FACT protocol and their clearing times were measured. After 1 mm of the brain slice clearing, the blood vessels containing auto-fluorescent red blood cells were imaged by a z-stack motorized epifluorescent microscope. The 3D structures of the brain vessels were reconstructed by Imaris software

Results: Auto-fluorescent blood vessels were 3D imaged by the FACT in mouse brain cortex. Clearing tissues of mice and rats were carried out by the FACT on the brain slices, spinal cord, heart, lung, adrenal gland, pancreas, liver, esophagus, duodenum, jejunum, ileum, skeletal muscle, bladder, ovary, and uterus

Conclusion: The FACT protocol can be used for the murine whole tissue clearing. We highlighted that the 3D imaging of cortex vasculature can be done without antibody staining of non-perfused brain tissue, rather by a simple auto- fluorescence

Effects of antidepressants on parameters, melondiadehyde, and diphenyl-2-picryl-hydrazyl levels in mice spermatozoa

Background: Prescribing antidepressant drugs is becoming common. These drugs are known to affect sexual functions

Objective: The study is aimed to assess the effects of amitriptyline and venlafaxine on sperm parameters and evaluate Malondialdehyde [MDA] and 1, 1-Diphenyl-2- picryl-hydrazyl values in BALB/ mice spermatozoa

Materials and Methods: Forty adult male BALB/c mice were separated into five groups. Group I [control] received distilled water; group II amitriptyline [4 mg/kg]; group III amitriptyline [4 mg/kg] +vitamin C [10 mg/kg]; group IV venlafaxine [2 mg/kg]; and group V received vitamin C [10 mg/kg] + venlafaxine [2 mg/kg]. All drugs were administered by oral gavage for 35 days. After excision of caudal epididymis, it was located in 1 mL Ham's F10 medium at 37 degree C for 15 min and then analysis of sperm parameters was performed. To examine lipid peroxidation and total antioxidant capacity, the MDA and 1, 1-Diphenyl-2-picryl-hydrazyl were measured, respectively

Results: The mean sperm parameters in the group treated with amitriptyline were significantly lower than in the other groups. MDA tests showed a significant difference between amitriptyline and control groups [p=0.007]

Conclusion: The results of this study showed that amitriptyline consumption can weaken sperm parameters, which can be attributed to the increased production of ROS and toxicity resulting from amitriptyline consumption. Moreover, venlafaxine improved sperm parameters in mice and the lipid peroxidation in this group did not change compared to the control group

Administration of high dose of methamphetamine has detrimental effects on sperm parameters and DNA integrity in mice

Background: methamphetamine [MA] was shown to have harmful effects on male reproductive system

Objective: to investigate probable effects of daily administration of MA on sperm parameters and chromatin/DNA integrity in mouse

Material and Methods: thirty-five NMRI male mice were divided into five groups including low, medium, and high dosage groups which were injected intraperitoneally with 4, 8 and 15 mg/kg/day for 35 days, respectively. Normal saline was injected in sham group and no medications were used in control group. Then, the mice were killed and caudal epididymis of each animal was cut and placed in Ham's F10 medium for sperm retrieval. To evaluate sperm chromatin abnormalities, the aniline blue, toluidine blue and chromomycine A3 were used. For sperm DNA integrity and apoptosis, the acridine orange, sperm chromatin dispersion, and TUNEL assay were applied. For sperm morphology, Papanicolaou staining was done

Results: normal morphology and progressive motility of spermatozoa decreased in medium and high dosage groups in comparison with the control group [p=0.035]. There was a significant increase in rate of aniline blue, toluidine blue, and chromomycine A3 positive spermatozoa in high dosage group. In a similar manner, there was an increase in rates of acridine orange, TUNEL and sperm chromatin dispersion positive sperm cells in high dosage group with respect to others

Conclusion: MA abuse in a dose-dependent manner could have detrimental effects on male reproductive indices including sperm parameters and sperm chromatin/DNA integrity in mice

Effect of Citrullus colocynthis hydro-alcoholic extract on hormonal and folliculogenesis process in estradiol valerate-induced PCOs rats model: An experimental study

Background: Citrullus colocynthis [CCT] is used as the anti-diabetic and antioxidant agent. Polycystic ovarian syndrome [PCOS] is a reproductive disorder which level of gonadotropins and sexual hormones are imbalanced

Objective: We evaluated the effect of CCT hydro-alcoholic extract on hormonal and folliculogenesis process in estradiol valerate-induced PCOs rats' model

Materials and Methods: 40 female adult Wistar rats divided into five groups [n=8 each: Group I [control] only injected by sesame oil as estradiol valerate solvent, group II [Sham] was orally received normal saline after estradiol valerate- induced polycystic ovarian syndrome [4 mg/rat estradiol valerate, intramuscularly], and three experimental groups, that after induction of PCOS within 60 days, received orally 50 mg/kg CCT extract [group III], 50mg/kg metformin [group IV], and CCT extract+ metformin [group V] for 20 days. The serum concentration level of luteinizing, testosterone and follicle stimulating hormones were measured using ELISA method and the serum concentration level of glucose were measured using the oxidative method [glucose meter]. Histological study of ovary tissue carried out by hematoxylin-eosin staining

Results: There was a significant reduction in luteinizing hormone and testosterone in III-V groups compared to Sham group, whereas follicle stimulating hormone in III-V groups was not significantly changed in comparison with Sham group. Histological investigations showed a significant increase in number of preantral and antral follicles and corpus luteum in the experimental groups compared to group II

Conclusion: Marked improvement in hormonal and histological symptoms of PCOS may be due to CCT effects hence, CCT can potentially be considered as an effective drug for treatment of PCOS

Effects of gamma radiation on fetal development in mice

Background: Many cancer patients receive radiotherapy which may lead to serious damages to the ovary storage and the matrix muscle state. Some of these patients may admit to infertility clinics for having pregnancy and on the other hand hormonal administration for superovulation induction is a routine procedure in assisted reproduction technology [ART] clinics

Objective: This study aimed to investigate fertility and fetuses of hormone treated super ovulated female mice who had received whole-body gamma irradiation before mating.Materials and Methods: Female mice were randomly categorized into a control group and 3 experimental groups including: Group I [Irradiation], Group II [Superovulation], and Group III [Superovulation and Irradiation]. In hormone treated groups, mice were injected with different doses of 59Tpregnant mare's serum gonadotropin59T [PMSG] followed with human chorionic gonadotropin [HCG]. Irradiation was done using a Co-60 gamma ray generator with doses of 2 and 4 Gy. Number of fetuses counted and the fetus's weight, head circumference, birth height, the number of live healthy fetuses, the number of fetuses with detected anomalies in the body, the sum of resorption and arrested fetuses were all recorded as outcome of treatments

Results: In the group I and group II, increased radiation and hormone dose led to a decrease in the number of survived fetuses [45 in 2 Gy vs. 29 in 4 Gy for irradiated group] as well as from 76 in 10 units into 48 in 15 units. In the group III, a higher dose of hormone in the presence of a 2 Gy irradiation boosted the slink rate; i.e. the number of aborted fetuses reached 21 cases while applying the dose of 15 Iu, whereas 6 cases of abortion were reported applying the hormone with a lower dose. Among different parameters studied, there was a significant difference in parameters of weight and height in the mouse fetuses [p=0.01]

Conclusion: The data indicated that use of ovarian stimulating hormones in mice that received pre mating gamma irradiation did not significantly increase the pregnancy rates

Survival assessment of mouse preimplantation embryos after exposure to cell phone radiation

Background: Using cellular phone has rapidly increased all over the world. Also, the concern on the possible health hazards of electromagnetic fields [EMF] induced from cell phones to reproduction has been growing in many countries. The aim of this study was to assess the consequences and effects of exposure to the cell phone radiation on the quality and survival rates of preimplantation embryos in mice

Methods: A total of 40 mice [20 females and 20 males], 6 weeks old and sexually mature BALB/c, were used for control and experimental groups. The ovary burses were removed and the zygotes were dissected in the morning after mating. Next, 2-cell embryos were divided into two groups of control [n=150] and experimental [n=150]. EMF [900-1800 MHz] was used for four days in experimental group for 30 min/day in culture at 37°C in a CO[2]incubator. The quality of embryos was recorded daily and the fluorescent staining was used for identification of viable blastocysts. All data were compared by Student's t-test and Mann-Whitney test [p<0.05]

Results: The rate of embryo survival to the blastocysts stage was similar in both groups. However, the percentage of dead embryos at the 2-cell stage was significantly higher in EMF-exposed group compared with controls [p=0.03]. Also, the loss of cell viability significantly increased in experimental blastocysts [p=0.002]

Conclusion: The normal embryonic development up to the blastocyst stage indicates that EMF-exposure commonly did not have adverse effect on embryo development in mice. But, it caused loss of blastocysts cell viability

Effect of prenatal methamphetamine administration during gestational days on mice

Methamphetamine [MA] is one of most common illicit drugs which were reported that nearly half of MA consumers are women. MA can cross through placenta and affects pregnancy and fetus development. Our aim was to evaluate effects of injected MA on crown-rump length, head and placental circumference, body weight, histological changes and apoptosis in fetus. Twenty-four NMRI pregnant mice were randomly divided into five groups. First, second and third groups were injected intraperitoneally 10 mg/kg/day MA during gestational days [GD]: GD1-7, GD8-14, and GD1-14, respectively. Forth group, as sham, was injected saline from GD1-14, and finally control which was received neither MA nor saline. On GD15 cervical dislocated pregnant mice, fetus and placenta were weighed and fetus crown-rump length was measured. Hematoxylin and Eosin staining and TUNEL assay were applied to assess histological changes and apoptosis respectively. Fetus body weight and crown-rump length showed significant decrease in third compared to first and second groups [p

Evaluation of embryo quality after concurrent use of ovarian stimulating hormones and gamma irradiation

Background: Radiotherapy has many side effects on fertilization in young women. Radiation can lead to ovarian failure in women who underwent abdomen or pelvic radiotherapy

Objective: This study helps us to investigate ovarian response of NMRI female mice to ovarian stimulating hormones [PMSG, HCG] after whole-body gamma irradiation

Materials and Methods: 45 pregnant mice were divided into two groups of control and experimental. The experimental group was classified into three sub-groups: Irradiation group [2 or 4Gy],Superovulation group [10 or 15IU],and superovulation and gamma-radiation group [2Gy and 10IU, 2Gy and 15IU, 4Gy and 10IU,4Gy and 15IU]. Female mice were killed and embryos were removed from oviduct .The number of embryos cells counted and the quality of them was evaluated in each group. Kruskal-Wallis test and Mann-Whitney test were used to analyze the data

Results: There was a significant difference in the number of 2-4 cells grade D embryos in 2Gy and 15IU group compared with control and 2Gy groups [p=0.01], and the number of embryos in 4Gy group was more than in 10IU and 15IU [p=0.03] and 2Gy and 15IU groups [p=0.01]. It was more significantly embryos in 4Gy and 15IU group compared to 2Gy and 15IU group [p=0.01].In addition There were no significant differences in the number of 2-4 cells grades A, B and C embryos and also number of 4-8 cells grades A, B and C, D embryos in groups

Conclusion: The concurrent use of ovulation stimulating hormones and gamma rays ameliorates this problem of drastic decrease in number of living embryos due to whole-body irradiation