Anatomical Sciences Journal
Year:2008 | Volume:6 | Issue:24|Papers Count:11

Purpose: The present study was designed to investigate the effect of hydrostatic pressure on cell viability, apoptosis induction, morphology and cell-substrate interactions of PC12 cells.Materials and Methods: PC12 as a neuronal cell line maintained in RPMI 1640 culture medium supplemented with 10% fetal bovine serum. PC12 cells were subjected to hydrostatic pressure. Experimental pressure condition was 100mmHg set above atmospheric pressure for 2 h. Controls were treated identically except for the application of pressure. Dye exclusion was used for viability assay, TUNEL staining was used for apoptosis detection. Cell area was assessed as morphometry and then cell adhesion, extension and migration were investigated. Results: Hydrostatic pressure had not changed viability of cells. It induced apoptosis in PC12 cells. In addition, hydrostatic pressure reduced cell area, adhesion, extension and migration ability of these cells (P<0.05). Conclusion: Hydrostatic pressure may induce apoptosis in PC12 cells as a result of inappropriate cell to substrate adhesion. Thus it is suggest that occurring apoptosis in these cells be an anoikis cell death induced by loss of attachment to the substrate.

Purpose: Dedifferentiation of the chondrocyte from rat articular cartilage with multiple subcultures and study of the redifferentiation potential of the cells into bone, cartilage and fat cell lineages.Materials and Methods: In this experimental study, chondrocytes from rat articular cartilage were isolated and expanded through several successive subcultures during which the expression levels of cartilage-specific genes including aggreacan and type II collagen were measured by using real-time PCR to determine the cell dedifferentiation (the time in which cartilage genes ceased their expression). Furthermore, during the culture period, the chondrocyte was examined morphologically by scanning electron microscopy (SEM). At the end, the dedifferentiated cells were subjected to osteogenic, adipogenic and chondrogenic culture condition to investigate whether or not they are able to redifferentaite into specialized progenies. Differentiation state was examined by specific staining and RT-PCR analysis.Results: Based on the findings by real time PCR, the expression levels of the both studied genes were high at passage 2 and dramatically decreased at passage 4. Aggreacan expression ceased at passage 10 and collagen II stopped expressing at passage 6. SEM images indicated the flattened morphology of the cells at early passages and the fibroblastic appearance at late passages. Differentiation examination revealed that the dedifferentiated cells were readily differentiated into bone, adipose and cartilage cell lineages. Conclusion: Considering all aspects together, this concluded that articular chondrocyte gradually lost their differentiated state during the long-term culture and changed into multipotent cells capable of differentiating into skeletal cell lineages.

Purpose: The aim of this study was to investigate whether demecolicne treatment of matured bovine oocytes adversely affects the process of in vitro fertilization and embryo development.Materials and Methods: Bovine Cumulus Oocyte Complexes (COC's) were matured in vitro and then were randomly allocated to two treatment groups of common concentrations of demecolicne (0.05 and 0.4 mg/ml for 30 min) and a control group. COC's were then fertilized and cultured in vitro for up to 9 days when the ratios of in vitro embryo development and the viability of the hatched blastocysts were assessed and compared with the control group (p<0.05).Results: The ratios of the cleavage and blastocyst formation of demecolicne treated groups (0.4 and 0.05 mg/ml) were 68.6, 63.5% and 23.3, 32.8%, which were not significantly different from the control group (73.3, 29.0%), respectively. The results of cell-viability were also not significantly different between the control vs. treatment groups.Conclusion: Since the overall indices of in vitro embryo development revealed no significant difference between the demecolicne treated compared to control bovine oocytes, it seems that demecolicne treatment of matured bovine oocytes may not compromise their potency for further in vitro development.

Purpose: To investigate the effect of different methods of synchronization on sheep granulosa cell cycle.Materials and Methods: Granulosa cells were aspirated from ovarian follicles and plated in a DMEM medium containing 15% FBS. Upon 70-80% confluency, the medium of the primarycultured as well as the passaged-5 cells were replaced with the medium containing either 0.5% FBS for 24, 48 and 72 h or 0.5 mg mimosine for 24 h. In the last group the cells were cultured in a base medium for further 4 days. In the present investigation, for each culture system, the cells were examined in terms of their cell cycle stage using flow cytometry. Moreover, the cultures were investigated with respect to their apoptotic as well as the proliferating cell contents by using Brdu labeling and TUNEL staining.Results: At primary as well as passaged-5 cultures subjected to serum starvation for 24 h, the frequency of G0/G1, proliferating as well as apoptotic cells were similar to those of control group. At culture with 48 and 72h serum starvation, the percentage of G0/G1 cells tended to increase significantly to 83% and 85% at primary culture and 89% and 90% at passage-5 culture respectively. Moreover, treating the cultures with mimosine caused the G0/G1 cell to increase. The percentages of apoptotic cells in cultures with either serum starvation (for 24 and 48 h) or with mimosine did not increase compared to those of control cultures. According to our results, 72 h after serum starvation, frequency of the apoptotic cells appeared to increase significantly.

Purpose: In this study specific antibody have been used for tracing type IV collagen basement membrane during renal tubules morphogenesis in mouse fetuses.Materials and Methods: 20 female Balb/C mice were selected randomly and were kept under normal condition, finding vaginal plug was assumed as day zero of pregnancy. 12 pregnant mice were scarified by cervical dislocation in one of gestational days 13-18 and their fetuses were fixed, serially sectioned and immunohistochemistry study for tracing of collagen type IV in BMG was carried out. The same processes were used for kidneys preparation on 5, 10, 15 and 20 postnatal days newborns of 2 mothers for each day.Results: Based on our data, Collagen IV showed weak reaction on day 14 of gestation in tubular BM. The amount of collagen increased continuously until next days of fetal life and primary of 5 days postnatal in BM. After this period, collagen IV reaction was not showed significant change in newborns.Conclusion: These results indicate that developmental changes in various nephron segments from most immature stages to most differentiated structures are dependent to the type IV collagen expression.

Purpose: Study of the effects of lead on the ultrastructure of uterine endometrium. Materials and Methods: In this study 40 female Balb/c mice with the age of one week were divided into two control and experimental groups. The experimental group were daily injected intraperitoneally 75 microgram per gram of lead nitrate dissolved in physiological serum for two weeks. The mice of control group were received only physiological serum. After the end of the injection period, the mice were anesthetized and dissected, then their uterus were removed and divided into small pieces and transferred into karnovsky solution, After ward they were processed for electron microscope and prepared semithin and ultrathin sections. morphometry on electron micrographs were carried out by point counting and also the number of stromal cells in 50 fields of light microscope were counted too, the data was analyzed with SPSS software and student's T-Test. Results: The endometrium epithelial cells in experimental group in comparison to control one had vacuoles inside the cytoplasm and volume fractions of nucleus to cell and mitochondria to cell were statistically different P<0.05 and also the mean number of the stromal cells had increased (P<0.05).Conclusion: The entrance of the lead to the body from living environment in different ways can have adverse effects on the uterine endometrium.

Assisted reproductive techniques (ART) are considered as the first line of treatment in overcoming infertility. The scope of success of these methods depends on several factors including integrity of sperm and oocyte. Obviously, paternal genomic content has an important effect on fertilization potential in infertile couples. Thus, sperm DNA damage is a genomic disorder that exists in different degree in infertile men. Since the first reports on sperm DNA integrity, this subject has become the focus of numerous studies. It has been reported that individuals with high percentage of sperm DNA damage have lower sperm parameters and, fertilization rate. Therefore, this review is based on our current understanding of mechanisms involved in sperm DNA damage and also will describe different procedures for evaluation of sperm DNA damage.

The knowledge of surgeon from anatomical variation can inhance the recovery process of patients. Vascular variation and their accidental cutting during surgical procedure can produce some problems specially in arterial variation. So the anatomical and surgical department must not neglect to prapere some program in order to learn these variations to the students. The antero-inferior abdominal wall of white 35 years old man has dissected. It was revealed that the obturator artery is an accessory branch of inferior epigastric artery at the medial side of deep inguinal ring. Subsequently it descend medially, crossing the free margin of lacunar ligament to reach the obturator canal. This variation is potentially dangerous in operation of femoral hernia.