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Objective: In vitro transplantation [IVT] of spermatogonial stem cells [SSCs] is one of the most recent methods in transplantation in recent decades. In this study, IVT and SSCs homing on seminiferous tubules of host testis in organ culture have been studied
Materials and Methods: In this experimental study, human SSCs were isolated and their identities were confirmed by tracking their promyelocytic leukemia zinc finger [PLZF] protein. These cells were transplanted to adult azoospermia mouse testes using two methods, namely, IVT and in vivo transplantation as transplantation groups, and testes without transplantation of cells were assigned in the control group. Then histomorphometric, immunohistochemical and molecular studies were done after 2 weeks
Results: After two weeks, histomorphometric studies revealed that the number of subsided spermatogonial cells [SCs] and the percentage of tubules with subsided SCs in IVT and in vivo groups were significantly more than those in the control group [P<0.05]. Immunohistochemical studies in the transplantation groups confirmed that the PLZF protein was expressed in the cells subsided on the seminiferous tubule. Quantitative reverse-transcription polymerase chain reaction [qRT-PCR] demonstrated that the PLZF gene expression was only positive in the transplantation groups, but it was not significantly different between the IVT group and the in vivo group [P>0.05]
Conclusion: Testicular tissue culture conditions after SSC transplantation can help these cells subside on the seminiferous tubule basement membrane
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Objective: re-vitrification of embryos immediately after thawing or after a culture period could be used to preserve the extra embryos after embryo transfer. This study aims to clarify the effect of re-vitrification on Bax and Bcl-2 gene expressions of compact and early blastocyst stage embryos
Materials and Methods: this experimental study was performed on mouse embryos. We collected 8-cell stage embryos [n=400] from female mature mice, 60-62 hoursafter injection of human chorionic gonadotropin [hCG]. The embryos were divided into 5 groups: fresh [n=80], vitrified at the 8-cell stage [n=80], vitrified at the blastocyst stage [n=80], vitrified at the 8-cell stage, and re-vitrified at the compact [n=80] and early blastocyst stages [n=80]. Embryos were vitrified by cryolock. We analyzed the developmental rates of the vitrified-warmed embryos with the chi-square test. Quantitative polymerase chain reaction [qPCR] data were analyzed with SPSS version 16 using one-way analysis of variance [ANOVA] followed by Tukey's post hoc test. P<0.05 were considered statistically significant
Results: the expanded blastocyst formation rate showed a significant difference in re-vitrified embryos compared with fresh embryos [P<0.05]. However, this result was similar between the two re-vitrified groups. Our data showed a significant difference in expression of the Bax and Bcl-2 genes between re-vitrified and fresh embryos [P<0.05]. Expressions of the Bax and Bcl-2 genes showed no significant difference between the two re-vitrified groups
Conclusion: based on our study, re-vitrification could affect developmental rate and expressions of the Bax and Bcl2 genes
ABSTRACT
Objective: low intensity ultrasound [continues and pulsed] is a form of energy. Spermatogonial stem cells [SSCs] are at the base of male fertility. This study investigated the effects of low intensity ultrasound stimulation [LIUS] and low intensity pulsed ultrasound stimulation [LIUPS] on the expression of germ cell-specific and pluripotency genes in SSCs in vitro
Materials and Methods: in this experimental study, isolated SSCs from neonatal male mice were cultured in Dulbecco's Modified Eagle's Medium [DMEM] with 10% fetal bovine serum [FBS]. In addition, to confirm identification of SSCs, PLZF protein was detected positively in SSCs derived colonies. SSCs were stimulated by LIUS and LIUPS for 5 days, followed by assessment of expression of integrin-alpha6 [Itga6] and beta1 [Itgbeta1], as two germ cell-specific genes, and Oct- 4, as a pluripotency gene, on day 21st by quantitive reverse transcriptase-polymerase chain reaction [qRT-PCR]. To investigate the proliferation rate and colonization of SSCs in different groups, counting whole number of the cells and colonies as well as analysis of the respective diameters were performed on days 7[th], 14[th] and 21[st]. Data was analyzed by ANOVA test
Results: LIUS and LIUPS treatment of mouse SSCs increased expression of Itga6 and Itgbeta1 genes in the experimental groups, compared to the control group [P<0.05], whereas there was no significant difference between the groups, regarding the expression of Oct-4 gene. These treatments maintained survival rate, while they increased proliferation rate and colonization of SSCs during the first week of culture. However, within the second week, proliferation rate and colonization were decreased in the experimental groups
Conclusion: these results suggested that LIUS and LIUPS treatment had good effect on SSCs proliferation and colonization, based on the gene-specific marker expression during 21 days culture in vitro
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Background: Sheep industry has taken steps toward transforming itself into a more efficient and competitive field. There are many varieties of sheep breeds in the world that each of them serves a useful purpose in the economies of different civilizations. Ghezel sheep is one of the Iranian important breeds that are raised for meat, milk and wool. Field of spermatogonial cell technologies provides tools for genetic improvement of sheep herd and multiple opportunities for research. Spermatogonial cells are the only stem cells capable of transmitting genetic information to future generations
Methods: This study was designed to extend the technique of isolation and in vitro proliferation of spermatogonial cells in Ghezel sheep
Results: Isolated cells were characterized further by using specific markers for type A spermatogonia, including PLZF. Also, sertoli cells were characterized by vimentin which is a specific marker for sertoli cells. After 10 days of co-culture, viability rates of the cells was above 94.7%, but after the freezing process the viability rates were 74 percent
Conclusion: In this study, a standard method for isolation and in vitro proliferation of spermatogonial stem cells in Ghezel sheep was developed
ABSTRACT
Background: Establishment of a standardized animal endometriosis model is necessary for evaluation of new drug effects and for explaining different ethological aspects of this disease. For this purpose, we need a model which has more similarity to human endometriosis
Objective: Our objective was to establish an autologous endometriosis mouse model based on endogenous estrogen level and analyze the influence of estrus cycle on the maintenance of endometriotic lesions
Materials and Methods: In this experimental study, endometriotic lesions were induced in 52 female NMRI mice by suturing uterine tissue samples to the abdominal wall. The transplantation was either performed at proestrus/estrus or at metestrus/diestrus cycles. Urine-soaked beddings from males and also male vasectomized mice were transferred to the cages to synchronize and maintenance of estrus cycle in female mice. The mice were sacrificed after different transplantation periods [2, 4, 6 or 8 wk]. The lesions size, macroscopic growth, model success rate, histological and immune-histochemical analyses were assessed at the end
Results: From a total of 200 tissue samples sutured into the peritoneal cavity, 83 endometriotic lesions were confirmed by histopathology [41.5%]. Model success rate for proestrus/estrus mice was 60.7% vs. 79.2% for metestrus/diestrus mice. The endometriotic lesions had similar growth in both groups. Number of caspase-3, Ki67-positive cells and CD31-positive micro vessels were also similar in endometriotic lesions of two groups
Conclusion: If we maintain the endogenous estrogen levels in mice, we can induce endometriosis mouse model in both proestrus/estrus and metestrus/diestrus cycle without any significant difference
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Background: Spermatogonial stem cells are the foundation of spermatogenesis and male fertility. So, their maintenance and culture are very important
Objective: In this study, we assessed protective effects of the Calligonum on in vitro viability and apoptotic and antiapoptotic genes expression of spermatogonial stem cells
Materials and Methods: After 24 hr of culture, the spermatogonial stem cells were treated with 30 MuM dose of H[2]O[2] and then 10 Mug/ml the Calligonum extract was added for 3 wks. Viability was assessed by Trypan blue, apoptosis using PI-Annexin and finally Bax, Bcl-2 and P53 genes expression by Real-Time Polymerase chain reaction
Results: After 3 wk of treatment, viability in the Calligonum extract+H[2]O[2] group was significantly higher than H2O2 group alone [p=0.001]. In the Calligonum extract+H2O2 group, apoptosis, as well as expression of apoptotic genes [Bax and P53], was significantly lower than the group treated with H[2]O[2] alone
Conclusion: The results of this study showed that 30 MuM H[2]O[2] increased apoptosis but decreased viability in spermatogonial stem cells. Calligonum has antioxidant properties that can reduce apoptosis, Bax and P53 expression and increase the viability and Bcl-2 expression
ABSTRACT
Background: In vitro spermatogenesis has a long research history beginning in the early 20th century. This organ culture method was therefore abandoned, and alternative cell culture methods were chosen by many researchers. Here, whether Tnp1, Tekt1, and Plzf, which play a crucial role in spermatogenesis, can be expressed during testis organ culture was assessed
Methods: Testes of 10 mouse pups were first removed, and the testis tissue was then separated into smaller pieces of seminiferous tubules. The size of the pieces was arbitrary; approximately 1 mg in weight or 1 mm3 in size when compacted. Afterwards, the testis tissue fragments [1-3] were transferred to the hexahedrons, incubated in a culture incubator and cultured for 12 weeks. Histological assessment and molecular evaluation were carried out at the end of the study
Results: The results showed that the expression of Tekt1 as a mitotic gene in mouse pups decreased significantly [p /= 0.05]. Based on histological study, different types of spermatocytes and postmeiotic stages of germ cells could not be detected
Conclusion: This kind of three-dimensional culture can induce expression of post-meiotic gene, Tnp1, but only at the molecular level and not beyond meiosis
ABSTRACT
Background: Polycystic ovary syndrome [PCO] is one of the most common reasons for infertility. Calligonum as a plant possess some of the important antioxidants that can decrease oxidative stress
Objective: The effects of treatment with Calligonum as an antioxidant on ovary tissue of a PCO mouse model
Materials and Methods: Thirty female NMRI mice were divided into three groups [n=10/each]: control, PCO, and Calligonum. We induced PCO model with single dose of Estradiol valerate [40 mg/kg]. Then Calligonum [20 mg/kg] was intraperitoneally injected weekly for two months. The level of oxidative stress and total antioxidant capacity was assessed in the ovarian tissue by flow cytometry and fluorescence recovery after photobleaching, respectively, and the histological study was conducted by the morphometric method and embryo development with in vitro fertilization
Results: The obtained results showed that estradiol valerate was able to increase oxidative stress within the ovary and causes ovarian cysts after two months. The cyst formation was decreased in Calligonum group compared to PCO group [p=0.001]. The percentage of pre-antral and antral follicles significantly decreased in Calligonum group compared to PCO group [p=0.001]. The oxidative stress decreased in Calligonum group significantly compared to PCO group [p=0.001]. Calligonum can significantly increase the total antioxidant capacity of ovarian tissue [p=0.001] as well as the percentage of in vitro fertilization compared to the PCO group
Conclusion: Calligonum could decrease ovary cyst in PCO model, and improve in vitro fertilization rate. Also, Calligonum extract as an antioxidant could decrease oxidative stress in PCO model
ABSTRACT
Background: Prenatal drug exposure, as a common public health concern, is associated with an increased risk of adverse effects on early embryo development
Objective: To investigate the in vitro development of - embryo from experimentally Kerack-addicted mice
Materials and Methods: Twenty-five female mice were studied in five groups: control, vehicle, and three experimental groups of Kerack-dependent mice [I, II, and III] which received different doses of Kerack for 14 days. After the establishment of addiction model [7 days], experimental groups I, II, and III were given Kerack intraperitoneally at the doses of 5, 35, and 70 mg/kg, twice a day for a period of 7 days, respectively. The vehicle group received normal saline and lemon juice whilst the control group just received water and food. Morulae were obtained through oviduct flashing. The survived embryos were cultured in T6+ 5mg/ml bovine serum albumin. The developmental rates up to hatched stage daily and embryo quality [differential staining and Tunnel staining] were also assessed
Results: The developmental potential of embryos obtained from the addicted mother was significantly decreased in comparison with control group. There was a significant reduction in the rate of blastocyst formation in the high dose Kerack dependent group. However, in addicted mice there was reduction in the total cell number [40.92% vs. 65.08% in control] and, inner cell mass percentage [17.17% vs. 26.15% in control] while apoptotic cells numbers were increased [7.17 vs. 1.46 in control] [p<0.05]
Conclusion: The Kerack addiction during pregnancy retards preimplantation development and induces apoptosis
Subject(s)
Animals, Laboratory , Substance-Related Disorders , In Vitro Techniques , Mice , ApoptosisABSTRACT
Background: nowadays, oocytes and embryos vitrification has become a routine technique. Based on clinical judgment, re-vitrification maybe required. But little is known about re-vitrification impact on genes expression
Objective: the impact of re-vitrification on apoptotic and implanting genes, Bax, Bcl-2 and ErbB4, at compaction stage embryos were evaluated in this study
Materials and Methods: in this experimental study, 8 cell embryos [n=240] were collected from female mature mice, 60-62 hr post HCG injection. The embryos were divided randomly to 3 groups included: fresh [n=80], vitrified at 8 cell stage [n=80], vitrified at 8 cell stage thawed and re-vitrified at compaction stage [n=80]. Embryos were vitrified by using cryolock, [open system] described by Kuwayama. Q-PCR was used to examine the expression of Bax, Bcl2 ErbB4 genes in derived blastocysts
Results: our result showed that expanded blastocyst rate was similar between vitrified and re-vitrified groups, while re-vitrified embryos showed significant decrease in expanded blastocyst rate comparing with fresh embryos [p=0.03]. In addition, significant difference was observed on apoptotic gene expression when comparing re-vitrified and fresh embryos [p=0.004], however expression of Bax and Bcl-2 [apoptotic] genes didn't demonstrate a significant difference between re-vitrified and vitrified groups. The expression rate of ErbB4, an implantation gene was decreased in re-vitrified embryos comparing with fresh embryos [p=0.003], but it was similar between re-vitrified and vitrified embryos
Conclusion: re-vitrification can alter the expression of Bax, Bcl-2 and ErbB4 genes and developmental rate of mouse embryos in compaction stage
ABSTRACT
Background: Niche cells, regulating Spermatogonial Stem Cells [SSCs] fate are believedto have a reciprocal communication with SSCs. The present study was conductedto evaluate the effect of SSC elimination on the gene expression of Glial cell line-Derived Neurotrophic Factor [GDNF], Fibroblast Growth Factor 2 [FGF2] and KitLigand [KITLG], which are the main growth factors regulating SSCs development andsecreted by niche cells, primarily Sertoli cells
Methods: Following isolation, bovine testicular cells were cultured for 12 days on extracellularmatrix-coated plates. In the germ cell-removed group, the SSCs were removedfrom the in vitro culture using differential plating; however, in the controlgroup, no intervention in the culture was performed. Colony formation of SSCs wasevaluated using an inverted microscope. The gene expression of growth factors andspermatogonia markers were assessed using quantitative real time PCR
Results: SSCs colonies were developed in the control group but they were rarely observedin the germ cell-removed group; moreover, the expression of spermatogoniamarkers was detected in the control group while it was not observed in the germ cellremovedgroup, substantiating the success of SSCs removal. The expression of Gdnfand Fgf2 was greater in the germ cell-removed than control group [p<0.05], whereasthe expression of Kitlg was lower in the germ cell-removed than control group [p<0.05]
Conclusion: In conclusion, the results revealed that niche cells respond to SSCs removalby upregulation of GDNF and FGF2, and downregulation of KITLG in order to stimulateself-renewal and arrest differentiation
ABSTRACT
Objective: Vitrification is a convenient, effective method for freezing and storing embryos. Under certain situations, such as an unsuitable endometrial environment, extra embryos can be re-vitrified for future use. There is inadequate data on the effects of revitrification on embryos, so we have evaluated the effects of re-vitrification on the development rate and expression of apoptotic and implantation genes
Methods: Female NMRI mice, ages six-eight weeks were super-ovulated with 7.5 IU PMSG and 7.5 IU hCG. Females were mated with males from the same strain and inspected for the presence of vaginal plugs the following morning. Females with the presence of vaginal plugs were considered to be pregnant and killed 62 h post hCG injection. Eight-cell embryos were flushed from their oviducts and subsequently divided into three experimental groups: fresh, vitrified-warmed 8-cell embryos, and re-vitrifiedwarmed blastocyst embryos. RNA was extracted and we used real-time PCR to evaluate expressions of Bax, Bcl-2, and ErbB4. Data was analyzed by the chi-square and ANOVA tests
Results: A significant difference existed in blastocyst formation rate, degeneration rate, and expressions of Bax, Bcl-2, and ErbB4 in re-vitrified embryos compared to fresh embryos
Conclusion: The vitrification and warming process did not affect the developmental rate and expressions of Bax, Bcl-2, and ErbB4 in the eight-cell stage embryos. However, we observed a change in development rate and expression rates of Bax, Bcl-2, and ErbB4 after re-vitrification in the early blastocyst stage
ABSTRACT
Objective: The goal was to evaluate the effect of LC on some indicators of embryo development and blastocyst quality including zona pellucid [ZP] thickness, the hatching of blastocysts and their cell numbers
Materials and Methods: Mouse embryos were randomly divided into five groups and incubated with different concentrations of LC [I; 0, II; 0.5, III; 1, IV; 2 and V; 4 mg/ml] from 2-cell to hatched blastocyst
The percentage of blastocysts and hatched blastocysts was calculated. Blastocysts ZP thickness was measured and the number of blastocyst cells was counted using Hoechst and propidium iodide [PI] staining
Results: The results showed concentration of 0.5 mg/ml of LC had an antioxidant effect as in this group, the percentage of blastocysts and hatched blactocysts [p=0.01], the ZP thickness [p=0.00] and the number of blastocyst inner cell mass were significantly more favorable than the control group [p=0.03]; and concentration of 4 mg/ml of LC had a toxic effect on embryo development and blastocyst quality [p=0.00]
Conclusion: The results suggest that LC may increase the number of blastocyst cells, which probably helps to expand the blastocyst and thinning of the ZP thickness and, therefore, creating a successful hatching for implantation
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Recently there is a focus on the antioxidants as adjuvant treatment of polycystic ovary syndrome [PCOS], the most endocrinopathy in reproductive age women. The aim of this review is answer to the question whether antioxidants are effective for managing of hormonal and metabolic problems in women with PCOS based on first degree evidences from Iran. A systematic review of clinical trials was done in Persian and international databases including PubMed, Scientific Information Database, Google Scholar, Iran Medex, and Magiran up to 2013. Keywords were including polycystic ovary syndrome, Iran, vitamin, antioxidant. From 440 potential studies found electronically, 11 studies; including 444 women in intervention and 390 women in control groups. Intervention in three studies was Calcium-vitamin D or calcitriol; in three studies was omega-3 fatty acids; in two studies was N-acetyl cysteine; in one study was folic acid; in one study was Zinc; and in one study was Soy. Finally, 11 studies that were relevant and met the inclusion criteria reviewed. There were 7 studies in English and 4 studies in Persian. We couldn't include all studies because all full texts were not accessible. The results showed that antioxidants and vitamins have positive effects on management of PCOS women. Although it seems more studies is necessary in this field
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Introduction: Nowadays, spermatogonial stem cells [SSCs] cultivation has been used by many researchers as an effective tool for infertility treatments. Oxidative conditions can be effective on cell proliferation and differentiation of these cells. So, the aim of this study was to establish oxidative stress model for antioxidant activity of some drugs investigation during SSCs in vitro culture
Methods: Neonatal NMRI male mice [3-5 day] were used for isolation of SSCs. The cell suspension was prepared by twice enzymatic digestion. The cell suspension contents were spermatogonial and sertoli cells and treated by different doses of H2O2 logarithmic concentrations from 0-100 and muM after 24 hours. To access the optimal dose, extra doses from 10-100 and muM was evaluated. After 2 hours of H2O2 treatment, viability was determined by Trypan blue assay. The data were analyzed using SPSS software and One-way ANOVA test
Results: Our data showed that spermatogonial stem cells colonies appeared after 4 days of isolation. These cells expressed OCT4 and PLZF proteins. Many of spermatogonial stem cells were removed after using higher doses of H2O2. The results showed that 30 and muM concentration of H2O2 could induce oxidative stress in spermatogonial stem cell during in vitro culture
Conclusion: According to this study, 30 and muM concentration of H2O2 can cause cell death lower than 50% of total number of cells and can increase oxidative stress in cultivation of SSCs. This model is a suitable tool for studying some new antioxidant drugs
ABSTRACT
Catsper proteins are responsible for entering Ca[2+] to the cell and play an important role in sperm motility and male fertility. Antioxidants are vital for sperm motility too. Escanbil [Calligonum] extract possess some of the important antioxidant like Catechin and Quercetin. Here we investigated the effects of Escanbil [Calligonum] extract on the sperm parameters and the expressing of Catsper gene in aging male mice. In this animal study, firstly, dose response was performed by using these three doses of Escanbil [Calligonum] [10, 30 and 50 mg/kg]. 5 mice in each group were considered and Intra Peritoneal injection was done for 5 weeks. the sperm parameters analyzed and Terminal deoxynucleotidyl transferase dUTP nick end labeling [TUNEL]staining was done. 30 mg/kg dose was considered as optimum dose. Secondly: fifteen aging male mice [11-13 months] were divided into three groups: control, sham and experiment. The experiments were injected Intra peritonealy with Escanbil [Calligonum] extract [30mg/kg] weekly for up to 5 weeks. The sham group was injected Intra Peritoneal [DMSO]. Sperm parameters were analyzed. Expression of Catsper genes was analyzed by Real time PCR. Our results showed that after Escanbil [Calligonum] treatment [30 mg/kg], the sperm parameters were improved in experimental group [p<0.05]. Our data showed that there was a statistical significance difference between the expressions of Catsper 2, 4 in aging experiment group comparison with aging control group [p<0.05]. We investigated that the Escanbil [Calligonum] extract [30 mg/kg] can improve sperm parameters and change the expression of Catsper genes in aging male mice. This herbal extract can be used as an antioxidant component for clinical usages
ABSTRACT
Spermatogonial stem cell [SSC] is a self-renewing population of male adult stem cell. SSCs have a differentiation potential which are similar to embryonic stem cells. These Embryonic stem like [ES-like] cells can be a potential source for pluripotent cells for stem cell-based therapy. This study presents an economical and simple co-culture system for pluripotent stem cells generation from neonatal mouse testis. Isolated testicular cells were cultured in DMEM/F12. Characteristics of the isolated cells and obtained ES-like cell were immune-cytochemically confirmed by examining the presence of PLZF, vimentin, Oct4 and Nanog protein. Expression of the pluripotency and germ-cell specific genes was analyzed by qPCR in derived ES-like colony and SSCs respectively. The experiment results indicated that our method of obtaining pluripotent ES-like cells from spermatogonial cells [SCs] is simpler than the described methods. ES-like cells were immunopositive for pluripotency markers. ES-like cell qPCR results indicated significant increase in pluripotency genes expression and significant decrease in germ cell-specific genes expression. The results indicated that ES-like cell with pluripotency characteristic were generated from freshly isolated spermatogonial cells. The pluripotent stem cells provide a cellular reservoir usable for regenerative medicine instead of embryonic stem cells.
ABSTRACT
It is hypothesized that stem cells have the capability to form tumors after transplantation. Spermatogonial stem cells have proliferation potency and colonization ability related to express pluripotency genes such as c-Myc. The primary aim of this study is to investigate tumorigenicity ability of these cells after in vitro cultivation and inoculation in athymic animals. Spermatogonial stem cells from 3-5 day-old neonatal mice testes [NMRI] were cultured following two-step enzymatic digestion. After one month of culturing the spermatogonial stem cells, the obtained colonies were identified by Oct4 and PLZF markers. Expressions of Nanog, Oct4 and c-Myc pluripotency genes were subsequently studied. We subcutaneously inoculated 5 x 106 cells into athymic mice and assessed tumor formation after 8 weeks. Mouse embryonic stem cells [CCE line] were used as the positive control. Generated tumors were measured by a caliper. The colonies expressed Oct4 and PLZF proteins. Ratio of pluripotency gene expressions in these cells compared to embryonic stem cells significantly decreased [P
ABSTRACT
Diabetic neuropathy leads to axonal transport abnormalities. However its mechanism and the beneficial effects of exercise on these abnormalities are not well documented. The present study aims to investigate KIF1B mRNA in spinal cord sensory neuron tissue of Wistar male rats with diabetic neuropathy following endurance training. We randomly assigned 12 male Wistar rats into three groups: diabetic trained, diabetic untrained and healthy control. Intraperitoneal injection of a STZ [streptozotocin] solution [45 mg/kg] was used to induce diabetes. At two weeks after STZ injections, the mechanical allodynia and thermal hyperalgesia tests demonstrated the presence of diabetic neuropathy. A moderate endurance training protocol was performed for a sixweek period. At 24 hours after the final training session, the rats were sacrified and the L4-L6 sensory neurons of the spinal cord tissue were removed. KIF1B mRNA expression was performed using real time-PCR. Diabetic neuropathy led to increased KIF1B gene expression in the diabetic untrained group compared with the intact control group [p=0.03]. Compared with the diabetic untrained group, training significantly decreased KIF1B gene expression [P<0.05] and blood glucose levels [P=0.0001] in the diabetic trained group. KIF1B mRNA up-regulation in sensory neurons of STZ-diabetic rats is a factor which can be involved in abnormal axonal transport. Endurance training as a nonpharmacotherapy strategy can modulate and return KIF1B to approximate normal levels.
ABSTRACT
This study presents an efficient, cost-effective method to improve proliferation and colonization of spermatogonial stem cells [SSCs] in vitro. Isolated SSCs from neonate mice were cultured in DMEM culture medium with 10% fetal bovine serum [FBS]. In the first phase of the study, the temperature was controlled by low intensity pulsed ultrasound stimulation [LIPUS] of the plate that contained the culture medium. In the next phase, SSCs were stimulated by LIPUS with 200 mW/cm[2] with 20% and 40% duty cycle for five days. Proliferation and colonization of SSCs were on the seventh day. LIPUS treatment of mouse SSCs increased the proliferation rate and colonization of SSCs in the experimental groups compared to the control group. Average proliferation rate in the 20% duty cycle group was 1.46 +/- 0.06, in the 40% duty cycle group it was 2.00 +/- 0.1 and for the control group, it was 1.26 +/- 0.06. The average number of colonies in the 20% duty cycle group was 24 +/- 7.7, whereas the 40% duty cycle group had 62 +/- 1.4 colonies and the control group had an average of 19 +/- 5.5 colonies. Average colony diameters were as follows: 186.6 +/- 2.07 micro m [20% duty cycle group], 185.3 +/- 4.4 micro m [40% duty cycle group] and 190.0 +/- 2.0 micro m [control group]. Our results showed a significant increase in proliferation rate and number of colonies in the experimental groups compared to the control group [P<0.05], whereas no significant differences were observed between groups in colony diameters. These results suggested that LIPUS treatment can be an efficient, cost effective method to improve proliferation and colonization of SSCs during in vitro culture.