1 17286158 4 398 Morphometry The Study on Expression of Mous Oocyte and Preimplantation Embryc Mct1 and Mct3 Genes in Vivo and in Vitro Esmaeil Pour Tahereh b Hoseini Ahmad Kazemi Bahram Chehreh Asa Fatemeh Kie Pitter b Cell & Molecular Research Center, Shaheed Beheshti Medical University, Tehran, Iran. 1 5 2005 3 1 1 9 26 12 2021 21 04 2005 Purpose: The aim of this study was to assay the profile of MCT1 & MCD in mouse unfertilized & fertilized oocytes and preimplantation embryos In vivo and In vitro. Materials and Methods: The presence of mRNAs encoding MCT1 & MCD3 were determined On unfertilized and fertilized oocytes, 2-cell, morulae, blastocyst and cultured embryos in plus glucose KSOM, minus glucose KSOM and pulse glucose KSOM by using RNeasy mini column kit. Fertilized oocytes and mentioned embryos were collected 24, 48, 72 & 96 hr. post-hCG injection respectively. In experiments requiring in vitro derived embryos, zygotes were collected and denuded at 18 hr. post-hCG. The culture medium used was KSOM modified to be either glucose-free (KSOM-G) or containing 1mM glucose. Embryos cultured in KSOM-G group were exposed to 27mM glucose at 65 hr. post-hCG for 2 hours (pulse G). Embryos of these three groups were collected at 90 hr. post-hCG injection. Transcripts encoding MCT1 & MCT3 were reversed transcribed in total 201 reaction mixture. PCR products were electrophorized on %2 agarose gel, stained by ethidium bromide and visualized by UV transiluminator. Density of the obtained DNA bands was determined by using Worklab & Excel soft wares. Results: MCT3 transcripts are not detected in mouse oocytes & embryos. MCT1 transcripts were present in oocytes, mentioned developmental stages and cultured embryos except KSOM-G group. DNA density is minimum in 2-C stage and maximum in blastocyst. Conclusion: It can be concluded that MCT1 gene which plays an important role in transport lactate & pyruvatein embryos, can be expressed in mouse oocyte and perimplantation embryos. The Presence of MCT1 mRNA in cultured embryos with glucose and exposed to glucose about 2- or 4-cell stage suggests that exposure of embryos to glucose during movement in oviduct is very important for expression of this gene and maybe is important for continuating transport of pyruvate and lactate across the cell membrane.
399 Morphometry Differentiation of Mouse Embryonic Stem Cells into Hematopoietic Cells Soleimani Masood g Atashi Amir Masoomi Mohammad Bahar Vand Hosein g Department of stem cells, Royan Institute, Tehran, Iran. 1 5 2005 3 1 11 18 26 12 2021 21 04 2005 Purpose: Differentiation of Mouse embryonicstem cells into Hematopoietic cells. Materials and Methods: In this study, we used EB formation system for Hematopoietic differentiation of mouse embryonic stem cell (Royan B1) in suspension culture. EBs cultured in medium with Hematopoietic inducer cytokines (SCF, TPO, GMCSF, IL3, Flt3 and EPO) .presence of hematopoietic differentiated cell assessed by RT-PCR and colony forming assay. Results: RT-PCR results showed the expression of two chain of mouse hemoglobin in differentiated cell. Also, colony forming assay confirmed results of RT-PCR. Colony of hematopoietic cells and benzidine positive cells that are representative of erythroid colonies were seen in semisolid media. Conclusion: The results of this study showed that cultivation of EBs in suspension medium with hematopoietic inducer cytokines is effective method for differentiation of embryonic stem cells into hematopoietic cells. 400 Morphometry The Campeare of the Effects Progesterone and Calcium Lonophore (A23187) on Mouse Sperm Parameters and Fertilization Rate Shah Verdi Abdol Hosein k Movahedin Mansoureh Reza Zadeh Valoujerdi Mojtaba Kazemi Saeed k Anatomy Department, Tarbiat Modarres University, Tehran, Iran. 1 5 2005 3 1 19 25 26 12 2021 21 04 2005 Purpose: The aim of this study to investigate the effect of progestrone and calcium ionophore (A23187) on mouse sperm parameters and fertilization rate. Materials and Methods: Csauda epididymidis were then obtained from NMRI male mice (10-12 wk old) and were divided to three groups: Control, adding progesterone (10µg/ml) and addingionophore (10mM) After sperm were prepared and incuhated in T6 medium in an atmosphere of 95% air, 5% Co2 and 37˚C sperm parameters were analized as WHO.The oocytes were also ohtained the same strain of female NRMI mice (7-10 wk old) and inseminated with sperm. Then rate of fertilization and embryo development were cheked daily during 120h culture period. Finally the result of each groups were analyzed and comparied by ANOVA, X2 and exact fishers test. Results: The results indicated that the main of grade three of sperm movement in progesterone group was increased significantly (P<0.05) compared with control groups, were as the value was not significantly different in calcium ionophore compare to control. The number of embryos derived from sperms that treated with calcium nophore significantly were higher than two others groups (P<0.05). Conclusion: The progesterone and calcium ionophore can increased the sperm motility, and fertilization rate in mouse. 401 Morphometry The Effects of Testosterone Treatment on Structure of Mouse Testes Morteza zadeh Meimandi Farhad Movahedin Mansoureh p Mokhtari Dizaji Manijeh p Department of Anatomy, Tarbiat Modares University, Tehran, Iran. 1 5 2005 3 1 27 37 26 12 2021 21 04 2005 Purpose: There is a great need for a safe male contraceptive method, which can provide an alternative to the female contraceptive. Efforts to develop new method for men have been made during recent decads, but the most reliable method is still sterlization by vasectomy. The ue of GnRH antagonist for fertility regulation requites androgen supplementation to offset the decrease in serum testosterone level. In the present study, administration of testosterone was studied in mouse testis as a animal model. For this purpose, the effects of 25 mg/mouse of testosterone enanthate (single dose per week) on the structure of testis were assessed. Materials and Methods: Testes from three individual mice for each control and experimental group were analyzed. The following groups were studied: control group that received 25 mg/weekly normal salin during 6 weeks, Experimental group that received 25 mg/weekly testosterone enanthate during 6 weeks. To study the reversibility of azoospermia two groups were studied three weeks without any injection after the last testosterone treatment. Sperm parameters (viability rate, motility rate and sperm count) and also histological morphometry were evaluated in all the groups. To do these, the adult male NMRI mice were killed by cervical dislocation and epidydymal sperm was obtained and viability, motility rate and sperm count were assessed. The left testis was chosen for morphometirc study and after fixation, processing, embedding in paraffin, sectioning and staming with Hematoxilin & Eosin, morphometirc measurement including tubular diameter, epithelium height, numbers of spermatogonia, spermatocyte and round spermatid cells were done. The data were analyzed using ANOV A and post hoc (TUKEY) tests. Results: The results showed that high dose of testosterone could induce azoospermia and normospermia was observed after finishing of injection. The same resuts obtained at histology level of testes. The results of the present study clearly show that proper dosage of testosterone that is critical in the induction of uniform long-term azoospermia is 25 mg/mouse weakly. This is first report where this dose of testosterone is sage and reversible in mouse. Conclusion: Reversible azoospermia or oligozoospermia can be induced by high doses of testosterone. Testosterone can decrease the numbers of spermatogonia, spermatocyte and spermatid cel. And as a result spermatozoa production diminished. 402 Morphometry Morphology and Ultrastructure of Mouse Polarized Endometrial Epithelial Cell Monolyer in Vitro Azad Bakht Mehri Reza Zadeh Valoujerdi Mojtaba Moula Seyyed Javad Anatomy Department, Tarbiat Modarres University, Tehran, Iran. 1 5 2005 3 1 39 48 26 12 2021 21 04 2005 Purpose: The objective for this study is to investigate the morphology and ultrastructure of mouse endometrial epithelial cell monolayer cultured on matrigel in dual-chambered system as an in vitro mouse endometrial epithelial cell culture model that mimics structural and functional properties of the endometrial epithelium in vivo. Materials and Methods: Mouse endometrial epithelial cells were enzymatically isolated using collagenase 0.25% and were seeded on Millipore filter insert coated with extracellular matrix Matrigel. The epithelial cells were studied microscopically by scanning electron microscopy. Tissue from uterine horn was also processed as control. Results: Light microscopy was showed that, the monolayer cultured on matrigel resemble a confluence layer. Scanning electron microscopy revealed a confluence epithelium with abundance large and smooth membrane projections as well as pinopods. Conclusion: Mouse endometrial epithelial cells in this culture model displayed characteristics near to the same cells in vivo. It can be suggested that this culture system may be provides a unique experimental model for basic research related to reproductive biology, endometrial pathology and basic cell biology issues. 403 Morphometry Effect of Electromagnetic Fields on the Male Mice Reproductive System Nik Ravesh Mohammad Reza Jalali Mahdi Anatomy Department, Medicine Faculty, Medical Sciences Mashhad University, Mashhad, Iran. 1 5 2005 3 1 49 55 26 12 2021 21 04 2005 Purpose: Previous studies have been shown that electromagnetic fields (EMF) can affect on functional activity of the female mice reproductive system. The aim of this study was to evaluated the effect of low frequency of EMF on histopathological changes of young male mice reproductive system. Materials and Methods: Eighteen male balb/c mice (6 week ages) were divided -at random into 3 groups of 6 animals each. Two experimental groups were exposed to a 50 Hz magnetic field at 0.1 or 0.5 mT. for 8 hours/day for 10 days and another group exposed to sham condition served as the control. After the end of the exposure, the mice were anesthetized and sacrificed by cervical dislocation. Their testis were removed for processing and serially section and histological staining methods for light microscopic study were done. Results: Quantitative analyses tubular germ cells showed a significantly decreasing number in the EMF exposure group 2 in comparison to experimental 1 and control (P<0.005). EMF exposure groups showed also an increasing internal diameter in somniferous tubules but external diameter in various groups was not significantly. Conclusion: Continuous exposure to low frequency of EMF may result in significant structural changes in male reproductive system and affect on organization of seminiferous tubules and spermatogenesis. 404 Morphometry A New Division of the Human Claustrum Basis on the Anatomical Landmarks and Morphological Findings Nam Avar Mohammad Reza Sadeghi Yoosef Haghir Hosein Anatomy Department, Medical Sciences Shiraz University, Shiraz, Iran. 1 5 2005 3 1 57 66 26 12 2021 21 04 2005 Purpose: The subdivision of claustrum into parts in some species exists in literature. Those are mainly based on a pattern of its connections with various cortical areas, method of staining, immunoreactivity of its neurons etc. The aim of this study was the division of the human claustrum into different parts, for first time, based on morphology, density, arrangement of claustral neurons as well as shape of the structure on coronal sections studied with Kluver-Barrera. Materials and Methods: Ten human brains (5 males, 5 females), who died of non-neurological causes and had no history of long term neurological illness were selected. The samples after fixation in formaldehyde and embedding in paraffin were coronally cut into 25-micrometer thick serial sections. Every 20th section was stained with Kluver-Barrera, with a random posi40n for the first section. The human claustrum was divided using anatomical landmark and morphological findings of neurons. Neuron density and total number of neuron and volume of each part were measured using stereo logical methods. Results: We have distinguished in the human claustrum three parts cap, dorsal and ventral in dorsa-ventral direction. The circular insular and rhinal sulci are upper and lower borders of dorsal part. Cap is the smallest and ventral part largest part. Neuron density decreases from cap towards ventral part. We have also found three parts anterior, central (middle) and posterior in antero-posterior direction. The central part is alongside of globus palliduas nucleus, whereas anterior and posterior are referred to parts of claustrum that lie in front and behind this nucleus, respectively. The central and posterior parts are the largest and smallest parts, respectively. Neuron density decreases from anterior towards posterior part, antero-posterior of human claustrum was about 28.06±5.25 mm. Conclusion: This study demonstrated that human claustrum consists of three anatomical parts in both dorso-ventral and antero-posterior directions. Each part has its specific connections, development, function and neuron density and morphology. 405 Morphometry The Effect of Laser Therapy of 30 Mw on Bone Repair: Histological Study Yousefi Diba Ali Asghar Mokhtari Dizaji Manizheh Ardeshir Larijani Mohammad Bagher Saleh Nia Mojdeh Torkaman Giti Medical Physics, Tarbiat Modarres University, Tehran, Iran. 1 5 2005 3 1 67 74 26 12 2021 21 04 2005 Purpose: The aim of the present study was to investigate the effect of Low Level Laser Therapy (LLLT) of 30mW with a Gallium Aluminum Arsenide (GaAlAs) diode laser device (830nm) on bone repair in the tibia of the rabbit, histological study. Materials and Methods: In this study, 24 male adult rabbits were randomly selected and anesthetized, then incision was made on the skin and deep fascia of medial aspect of the tibia of each rabbit. By using drill (3mm diameter), the medial of the tibia was drilled. The animals were randomly classified to three groups: control, sham and treatment. Irradiation was started 24 h after surgery and was applied daily in 5 sites of bone with energy density 126 J/Cm2. For evaluation of repair, the animals of each group were sacrificed on the 7th and 14th days after the irradiation. The samples were collected from repaired tissue in order to histological evaluation and compartment between groups. Results: The results show that in the 7-dayssubgroup regarding the osteogenesis no difference was observed among the control, sham and treatment groups, but osteogenesis areas in the 7-days treatment subgroup, in all area of the defect site, were observed to be more active than the control and sham groups. In 14-day treatment group defect sites were completed by spongy bone while in other groups no change was observed. Conclusion: It was concluded that in the process of bone fracture repair, photochemical interaction of laser had a effect on osteogenesis trend and in the 14th day treatment subgroup, this process is more effective than others.