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PubMed2004

Ultrastructure of spermiogenesis and spermatozoon of Leptorhynchoides plagicephalus (Acanthocephala, Palaeacanthocephala), a parasite of the sturgeon Acipenser naccarii (Osteichthyes, Acipenseriformes).

This paper describes the ultrastructure of spermiogenesis and the spermatozoon of Leptorhynchoides plagicephalus, an acanthocephalan parasite of the sturgeon Acipenser naccarii, a species which is under the threat of extinction. At the beginning, spermiogenesis in L. plagicephalus is characterized by the presence of a single centriole in the early spermatid. This centriole generates a flagellum with a 9+0 pattern. Another ultrastructural feature observed during the spermiogenesis of L. plagicephalus is the condensation of chromatin to form an "intranuclear wall". The mature spermatozoon of L. plagicephalus presents a reversed anatomy, as observed in other species of the Acanthocephala. The spermatozoon is divided into two parts: an axoneme and a nucleocytoplasmic derivative. The pattern of spermiogenesis and the ultrastructural organization of the spermatozoon of L. plagicephalus are compared with information available on other acanthocephalan species. The appearance of an "intranuclear wall" observed during the present study represents the first record within the Acanthocephala and is unknown from other animal taxa.

Acanthocephala↗
PubMed2004

The effect of topoisomerase II inhibitors on the kinetoplast ultrastructure.

Topoisomerases from trypanosomatids play key functions in the replication and organization of kinetoplast DNA (kDNA). Hence, they are considered as potential targets for anti-parasite drugs. In this paper, the effect of topoisomerase II inhibitors, such as nalidixic acid, novobiocin and etoposide, on the ultrastructure of trypanosomatids that present distinct kDNA arrangements was evaluated. Prokaryotic topoisomerase II inhibitors were more effective on growth arrest and ultrastructure changes than etoposide, a eukaryotic topoisomerase II inhibitor. With the exception of novobiocin, drug concentrations which inhibited cell proliferation also promoted kinetoplast ultrastructure alterations, including the redistribution of topoisomerase II. The data reinforce the concept that prokaryotic topoisomerase II inhibitors may offer greater selectivity in drug therapy of trypanosomatid infections.

Animals↗
PubMed2004

Ultrastructure of the spermatozoon of the anoplocephalid cestode Gallegoides arfaai (Mobedi and Ghadirian, 1977) Tenora and Mas-Coma, 1978, an intestinal parasite of the wood mouse (Apodemus sylvaticus Linnaeus, 1758).

The present work involves an ultrastructural study of the mature spermatozoon of the anoplocephalid cestode Gallegoides arfaai (Mobedi and Ghadirian, 1977) Tenora and Mas-Coma, 1978, obtained from the small intestine of naturally infected wood mice, Apodemus sylvaticus Linnaeus, 1758 (Rodentia, Muridae). The mature spermatozoon of G. arfaai is a filiform cell, tapered at both ends and lacking mitochondria. It is characterized by the presence of a 1,000-nm-long apical cone and two 140-nm-thick crest-like bodies in its anterior extremity. The axoneme, of the 9 + '1' trepaxonematan pattern, lacks a periaxonemal sheath, and disorganization occurs at the level of the nuclear region of the sperm cell. The cortical microtubules form two to four fields according to the different regions of the male gamete. They are twisted at an angle of about 35 degrees , becoming parallel towards the posterior extremity of spermatozoon. The nucleus, spiralled around the axoneme, shows an irregular shape in both longitudinal and cross-sections. Numerous electron-dense granules were observed, which transform into an electron-dense material in the posterior extremity of the cell. Moreover, we describe for the first time the total length of the anterior region of sperm containing the helical crest-like bodies. This anterior extremity measures around 15 mum and presents two helical crest-like bodies of different lengths that describe 13-14 turns around the sperm body. Our ultrastructural results on the G. arfaai spermatozoon are compared with the ultrastructural organization of the spermatozoa of other previously studied species, with particular emphasis on the anoplocephalids.

Animals↗
PubMed1997

The effect of high n-3 fatty acids diets on the ultrastructural development of Eimeria tenella.

A study of development of Eimeria tenella in chickens fed high n-3 fatty acids (n-3FA) diets showed ultrastructural degeneration of both asexual and sexual parasite stages. Abnormal shedding of asexual and sexual parasite developmental stages into the cecal lumen was also observed. Ultrastructural degeneration was characterized by cytoplasmic vacuolization, chromatin condensation within the nucleus, a lack of parasitophorous vacuole delineation, and, in some cases, a complete loss of parasite ultrastructural organization. The results of this study indicate that diets high in n-3FA may be useful in the control of avian coccidia.

Animal Feed↗
PubMed1999

Ultrastructural study of the papillae and presumed sensory receptors in the scolex of the Gymnorhynchus gigas plerocercoid (Cestoda: Trypanorhyncha).

The ultrastructure of the papillae and presumed sensory receptors in the tegument of the scolex of Gymnorhynchus gigas plerocercoid were investigated by scanning and transmission electron microscopy. Four distinct types of putative sense receptor (including three uniciliated and one nonciliated) are described for the first time in G. gigas, and this investigation is the first detailed ultrastructure study of tegumental receptors carried out in trypanorhynch cestodes. Microtriches arranged in clusters with a dome-shaped pattern seemed to be papillae that contained a ciliated sensory receptor (type I). The density of these papillae was greater in the center of the bothridial adherent surface near the tentacle orifice than in the lateral margins of the bothridia or in the pars post-bothridialis. The type I receptor is characterized by a long cilium anchored in the nerve bulb by a dense basal body but lacks rootlets. The bulb contains one or two electron-dense collars, numerous electron-lucent neurovesicles, and some mitochondria. The type II receptor presents a short cilium retracted into an invagination of the tegument that arises from the basal body. The bulb contains one electron-dense collar and numerous electron-lucent vesicles, but the rootlets are absent. The type III receptor is also a ciliated receptor embedded in a bulb wider than that of types I and II, and it differs from type II in that it possesses two electron-dense collars and small rootlets associated with the basal body. No electron-lucent vesicle was found in the bulb of this type III receptor. The type IV receptor is a nonciliated receptor localized under the surface with no contact to the outside. It consists of a flattened bulb that contains two electron-dense collars and striated rootlets associated with a band of microfilaments. A comparison of the ultrastructural features of receptors in different cestodes is presented.

Animals↗
PubMed2001

Ultrastructural damage of Trypanosoma cruzi epimastigotes exposed to decomplemented immune sera.

The susceptibility of Trypanosoma cruzi epimastigotes to lysis by normal or immune sera in a complement-dependent reaction has been reported, but the effects induced directly by immune serum depleted of complement remain unstudied. The aim of this work was to study the ultrastructural alterations induced in T. cruzi epimastigotes by immune mouse or rabbit sera with or without complement. A local isolate of T. cruzi (Queretaro) was used in all experiments. Immune sera were raised in both mouse and rabbit by immunization with T. cruzi epimastigote antigens. Light microscopy showed intense agglutination of epimastigotes when incubated with decomplemented mouse or rabbit immune sera. A distinctive ultrastructural feature of this agglutination pattern was the fusion of plasma membranes and a pattern of intercrossing between subpellicular microtubules. Agglutination was associated with fragmentation of nuclear membranes and swelling of cytoplasm, Golgi cisternae, endoplasmic reticulum, mitochondria and kinetoplast membranes. Agglutinated parasites also incorporated trypan blue stain. Results of [3H]-thymidine incorporation confirmed that epimastigotes exposed to specific antibodies in the absence of complement were incapable of proliferating. Ultrastructural changes observed in epimastigote micrographs incubated with decomplemented immune mouse sera were statistically significant (P<0.001) when compared with results obtained from images after incubation with decomplemented normal mouse sera.

Animals↗
PubMed2002

Nitric oxide level regulates the embryonic development of the pond snail Lymnaea stagnalis: pharmacological, behavioral, and ultrastructural studies.

On the basis of the distribution of NADPH-diaphorase (NADPH-d) activity, we have previously suggested a role for nitric oxide (NO) in the development of Lymnaea stagnalis. In the present study, the long-term effects of NO donors (sodium nitroprusside, S-nitroso-N-acetyl-penicillamine) and nitric oxide synthase (NOS) inhibitors (nitro-L-arginine methyl-ester [L-NAME], N(G)-nitro-L-arginine [L-NOARG]) were tested on the survival, length of embryonic (intracapsular) life, locomotion (gliding), heartbeat activity and feeding behavior, as well as on the ultrastructure of the developing ganglia in the embryonic Lymnaea. No effect of any of the substances applied can be observed under 10(-5) M concentration, whereas at 10(-3) M concentration both kinds of treatment proved to be toxic. Between 10(-5) M and 10(-3) M concentrations the effects are reversible. At 10(-4) M concentration, NO donors slightly increase the frequency of gliding and heartbeat of E70% embryos, and evoke a more than twofold enhancement of the feeding activity, i.e., the frequency of radula protrusions in the E90% embryonic stage. In contrast, NOS inhibitors at 10(-4) M concentration strongly inhibit the locomotion and heartbeat of E70% embryos, and the feeding of E90% embryos. Under 10(-3) M concentration, L-arginine diminishes the effect of NOARG, whereas the D-isomer of NAME has little or no significant effect. Neither type of treatment alters the course of gangliogenesis, and the light-microscopic appearance of neurons also remains unaffected. Ultrastructural analysis of the central nervous system of E90% embryos treated with 10(-4) M NOS inhibitors revealed a significant reduction of the glycogen granule content and accumulation of lipid droplets in a number of the neuronal perikarya, as well as the occurrence of disintegrated mitochondria in axonal profiles. The effect of 10(-4) M NO donors is mainly characterized by the increased number of lysosomes, disintegrated mitochondria and degenerating axonal profiles. The present findings suggest that NO is involved in the regulation of different behaviors and physiological functions, such as feeding activity, locomotion and heartbeat, during the embryonic development of Lymnaea. Changes observed in neuronal ultrastructure in ganglia seem to indicate NOergic regulatory processes at the central level.

Animals↗
PubMed2004

Ultrastructural changes in cirrhotic and noncirrhotic patients due to hepatectomy.

BACKGROUND/PURPOSE: Alterations at the ultrastructural level can be identified prior to histological change in the early phase of irreversible cell damage. The aim of this investigation was to compare the ultrastructural changes in cirrhotic and noncirrhotic liver in response to ischemic and reperfusion injury due to hepatectomy. METHODS: Hepatic resections using the same technique were performed in cirrhotic and noncirrhotic patients. Three biopsy specimens (Tru cut) from each patient, in the unresected part of the liver, were studied by transmission electron microscopy: immediately after laparotomy, before releasing of the porta hepatis clamp (ischemic phase), and 30-45 min after reperfusion. RESULTS: All patients did well after surgery, except for 1 cirrhotic patient who died of liver failure. There were no significant differences in operative time, blood loss, and inflow occlusion times in any of the 15 patients. We found that morphological changes were the same in the 10 non-cirrhotic and 4 cirrhotic patients. Changes during the ischemic phase included nuclear membrane deformity, focal chromatin condensation at the nuclear margin, and swelling of both mitochondria and endoplasmic reticulum. In the reperfusion phase, there were early irreversible changes in the nuclei of some hepatocytes and intramitochondrial particles and increased vacuolization in cytoplasm. Endothelial cells, Kupffer cells, bile canaliculi, and Ito cells were not affected in either the ischemic or the reperfusion phase. However, in the 1 cirrhotic patient who died of liver failure, there were marked swelling and dilated cristae in mitochondria during the ischemic phase and deformity of Ito cells during the reperfusion phase. CONCLUSIONS: In this, the first report of ultrastructural changes due to hepatectomy in cirrhotic patients, we found that the changes were the same as those in non-cirrhotic patients, except for the one cirrhotic patient who had postoperative liver failure.

Adult↗
PubMed2004

Ultrastructural changes in the contralateral lung tissue following unilateral lung ischemia: an experimental study in rabbits.

PURPOSE: To investigate the acute ultrastructural changes that may occur in the contralateral nonischemic lung tissue after unilateral ischemia of a lung in a rabbit model. METHODS: The animals were divided into three main groups of eight; namely, a 2-h procedure group, a 4-h procedure group, and an 8-h procedure group. Each of these groups was further divided into two subgroups of four rabbits each; namely, a control group, given a sham operation without any ischemic insult, and an ischemia group, in which the main pulmonary arteries, the pulmonary veins, and the main bronchi of the left lungs were ligated after thoracotomy. Tissue samples were taken from the left and right lungs to examine the ultrastructural changes after 2, 4, and 8 h of ischemia. Each sample was given a semiquantitative histological injury score. Statistical analysis was done by the Mann-Whitney U-test. RESULTS: Contralateral ultrastructural damage, evident by heterochromatin in the nuclei, mitochondrial degeneration, cisternal widening of the endoplasmic reticulum, increased lipid droplets, and lysosomes, was determined by electron microscopy after unilateral lung ischemia. The contralateral lung injury was significantly correlated with the duration of ischemia. CONCLUSIONS: Unilateral lung ischemia affected the bilateral lungs in a rabbit model. Therefore, in operations such as single-lung transplantation, pulmonectomy, or lobectomy, if the procedure is unnecessarily prolonged, the contralateral lung may be damaged, which could seriously affect the prognosis of the patient.

Animals↗
PubMed1998

Ultrastructural and hormonal changes in the contralateral adrenal gland in unilateral adrenal gland ischemia: an experimental study in rats.

While it is well known that unilateral tissue ischemia may result in contralateral damage in some paired organs, there is no universally accepted mechanism to explain why these contralateral changes occur. The aim of the present study was to investigate the ultrastructural and hormonal changes that occur in the contralateral nonischemic adrenal gland after unilateral ischemia of an adrenal gland in a rat model. The animals were divided into four groups of four rats each; namely, a control group which received a sham operation without any ischemic insult, a 2-h ischemic group, a 4-h ischemic group, and an 8-h ischemic group. The left adrenal blood vessels were ligated in all ischemia groups and blood samples were taken for hormonal study 2, 4, and 8 h later, after which bilateral adrenalectomy was performed to determine the ultrastructural changes. The plasma concentrations of aldosterone and cortisol were determined by radioimmunoassays. There was an increase in both aldosterone and cortisol levels related to the duration of the ischemia, but the differences among the groups were not statistically significant. Contralateral ultrastructural damage such as heterochromatin in nuclei, mitochondrial degeneration, endoplasmic reticulum cisternal widening, increased lipid droplets, and lysosomes, were demonstrated electron-microscopically after unilateral adrenal ischemia.

Adrenal Glands↗
PubMed2005

Blockade of sodium channels by phenytoin protects ultrastructure and attenuates lipid peroxidation in experimental spinal cord injury.

Spinal cord injury (SCI) involves a series of pathological events. Abnormal sodium influx has been implicated as one of the key events in the pathophysiology of the SCI. Pharmacological blockade of sodium channels can reduce secondary injury and increase recovery from trauma. The aim of the present study was to show the neuroprotective effect of phenytoin, a sodium channel blocker, after experimental SCI. Control and laminectomy-only groups were not injured. 50 g-cm weight drop injury was produced in the trauma group. In the treatment groups, methylprednisolone (30 mg/kg) and phenytoin (1 mg/kg, 10 mg/kg, or 30 mg/kg) were given intraperitoneally immediately after injury. Malondialdehyde (MDA) levels in the spinal cord samples were examined for lipid peroxidation. Spinal cord ultrastructure was evaluated and grading system was used for quantitative evaluation. Trauma increased tissue MDA levels. Treatment with methylprednisolone and phenytoin decreased MDA levels compared to trauma in all doses. Significant ultrastructural neuroprotection was observed with 30 mg/kg of phenytoin treatment according to general neural score. This ultrastructural neuroprotection of phenytoin was not different from methylprednisolone. Phenytoin appears to protect spinal cord against injury by decreasing lipid peroxidation and lessening neuronal damage associated with SCI in rats.

Animals↗
PubMed2005

Characteristics of histopathological and ultrastructural features of placental villi in pregnant Nepalese women.

The placenta is an important functional unit for gas transfer between mother and fetus. The placental membrane, consisting of trophoblast layer interposed between maternal and fetal blood, plays an active role for intensity of respiration, but no morphological evidence has been documented. Until now, it has been reported that fetal growth retardation and increased fetal mortality rate usually could be seen at high altitude. In an attempt to find the cause of high perinatal mortality rate in Nepal, this study was undertaken to examine pathologically about 1000 Himalayan placentas obtained in Nepal and Tibet since 1977, and the results were compared with those of 5500 Japanese placentas at Saitama Medical School since 1990. In this study, characteristics of ultrastructural features of the Nepalese placental villi investigated in recent years are reported. (1) The gross characteristics of placental pathology in the Himalayan group were represented by marked subchorionic fibrin deposits and increased chorionic cysts in contrast to low incidence of intervillous thrombosis compared with those of the Japanese group. (2) As characteristics of histological findings of the placental villi between Himalayan and Japanese groups, the incidence of chorangiosis and chorangioma in the Himalayan group was significantly higher than that in the Japanese group. (3) Accompanying an increase of vasculosyncytial membrane (VSM) in the villi, thickness and separation of basement membrane of the syncytium in addition to increased apoptosis of syncytial cell nuclei were recognized. (4) As characteristic ultrastructural features of chorionic villi of Nepalese placentas, an increase of mitochondria and cystic formation of rough endoplasmic reticulum (rER), in addition to appearance of lamellar bodies similar to alveolar epithelial type II cell in organellae of the syncytium, were observed. These ultrastructural changes of the placental villous capillaries may be ascribed to hypervascularization caused by the chronic hypoxic state. It is, therefore, presumed that trophoblast cells may play an important role for gas transfer mechanism under such a hypoxic state at high altitude.

Adult↗
PubMed2006

An ultrastructural study on the ligamentum flavum of the cervical spine in patients with ossification of the posterior longitudinal ligament.

Some histological analyses of the ossification of the posterior longitudinal ligament (OPLL) have been reported, but no ultrastructural studies of the ligamentum flavum (LF) in patients with OPLL have been published to date. To understand the pathology of the ossification of the spinal ligament, we examined, by electron microscopy, ultrastructural changes in the LF in cases of OPLL and made a comparison with the LF in cervical spondylotic myelopathy (CSM). Subjects were three men and two women with cervical OPLL who underwent longitudinal spinous process-splitting laminoplasty. During surgery, a small piece of the LF was collected from C2-C3 to C7-T1 and was then analyzed by light and electron microscopy. We observed atrophic elastic bundles with a two-layer structure and disarrangement, a partially torn area, the disappearance of microfibrils, and an enlarged interstitium with an irregular alignment of collagen fibrils. We observed some properties of a cell preceding its death: the initial phase may be the disappearance of the plasma-membrane, followed by the scattering of many organellae around its degenerated nucleus. Finally, many extracellular plasma membrane-invested particles that resemble matrix vesicles remain there without phagocytosis. These results suggest that ultrastructural abnormalities exist in the spinal ligament in cases of ossification of the spinal ligament.

Aged↗
PubMed2001

The ultrastructural and immunohistochemical heterogeneity of CD-30-positive neoplasms: so-called anaplastic large cell Ki-1 lymphomas.

Anaplastic large cell lymphoma (ALCL), also referred to as Ki-1 lymphomas, was first recognized as an entity with characteristic light microscopic appearance in 1985. This tumor is composed of variably cohesive cells, often with large, markedly atypical, and multinucleated cellular forms. The recognition of ALCL resulted from the development of a monoclonal antibody in Kiel, Germany, named Ki-1, which was initially believed to be a putative marker for Reed-Sternberg cells. This antibody was later found to be specific against the epitope CD-30. Attempts to create strict criteria to preserve this neoplasm as a specific entity have undergone evolution. However, it is now clear that included in this group are a variety of pleomorphic neoplasms with CD-30 immunoreactivity. Some of these neoplasms are nonlymphoid and show marked heterogeneity in their immunohistochemical and ultrastructural profiles. This article aims to highlight the ultrastructural spectrum of neoplasms exhibiting CD-30 positivity that are within the spectrum of ALCL. It remains to be determined if there are subgroups of these CD-30-positive neoplasms that can be segregated on the basis of ultrastructural and immunohistochemical criteria with corresponding clinical correlates that may impact on their management, treatment, and prognosis. We review here the heterogeneity of CD-30-positive neoplasms (so-called anaplastic large cell Ki-1 lymphomas).

Abdominal Neoplasms↗
PubMed2006

Etiology of inguinal hernia: ultrastructure of rectus sheath revisited.

In the last decade, in the search for abdominal-wall hernia etiology, attention has been brought to alterations in the connective tissue ultrastructure as the probable etiological factor. These may cause weakening of connective tissue, which in turn may form ground for hernia formation. To investigate this hypothesis in depth, we compared the ultrastructure of the connective tissue in hernia patients and the control group. The study group consisted of five patients with primary inguinal hernia (Nyhus II = 4, Nyhus IIIa = 1). Another five patients posted for emergency appendectomy created the control group. Tissue specimens, harvested intraoperatively from the rectus muscle sheath (RAMS) and fixed in 4% glutaraldehyde, underwent staining by the Masson, H-E and methylene blue techniques and were assessed by microscopy (light and scanning electron). The examinations showed significant differences in the rectus sheath ultrastructure. They included altered architecture, placement and quantity of collagen and elastic fibers, differences in the caliber of individual fibers and disrupted ground matter-to-fiber ratio. In patients with hernias, chaotic arrangement of collagen fibers was seen, as well as their thinning and a decrease in the general amount of elastic fibers, replaced by ground matter. Our research has shown significant differences in the structure of the RAMS between patients with hernias and healthy individuals. This supports the theory linking connective tissue alterations with the etiology of hernia, and stating that these alterations include connective tissue at locations distant from the hernia site as well, as the rectus sheath itself does not form a hernial defect.

Adolescent↗
PubMed2006

Ultrastructural characterization of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid-induced cell death in embryonic dopaminergic neurons.

Developing neuronal populations undergo significant attrition by natural cell death. Dopaminergic neurons in the substantia nigra pars compacta undergo apoptosis during synaptogenesis. Following this time window, destruction of the anatomic target of dopaminergic neurons results in dopaminergic cell death but the morphology is no longer apoptotic. We describe ultrastructural changes that appear unique to dying embryonic dopaminergic neurons. In primary cultures of mesencephalon, death of dopaminergic neurons is triggered by activation of glutamate receptors sensitive to alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), and differs ultrastructurally from both neuronal apoptosis or typical excitotoxicity. AMPA causes morphological changes selectively in dopaminergic neurons, without affecting other neurons in the same culture dishes. Two hours after the onset of treatment swelling of Golgi complexes is apparent. At 3 h, dopaminergic neurons display loss of membrane asymmetry (coinciding with commitment to die), as well as nuclear membrane invagination, irregular aggregation of chromatin, and mitochondrial swelling. Nuclear changes continue to worsen until loss of cytoplasmic structures and cell death begins to occur after 12 h. These changes are different from those described in neurons undergoing either apoptosis or excitotoxic death, but are similar to ultrastructural changes observed in spontaneous death of dopaminergic neurons in the natural mutant weaver mouse.

Animals↗
PubMed2005

Immunophenotypic and ultrastructural validation of a new human glioblastoma cell line.

1. A human glioma cell line, NG97, was established by Grippo et al. in 2001 from tissue obtained from a grade III astrocytoma (WHO, 2000). In this first study, the cell line grew as two morphologically distinct subpopulations: dendritic/spindle cells and small round cells. The injection of NG97 cells into nude mice induced an aggressive tumor characterized by: severe cytological atypia, vascular proliferation and pseudopalisading necrosis (glioblastoma multiforme features). 2. The purpose of the present study was to characterize the immunophenotype and ultrastructural aspects of this cell line, using the parental tumor, cultured cells and the xenotransplant, in order to assess its glial nature and possible divergent differentiation. 3. NG97 cells and xenotransplant expressed the main neuroglial markers (GFAP, S-100 protein, NSE and Leu-7) and showed no aberrant expression of other histogenetic markers. GFAP was similarly expressed in the parental tumor and in the cells in culture, but decreased in the xenotransplant. NSE expression was reduced in NG97 cells, but substantially recovered in the xenotransplant. This variability in expression of GFAP and NSE was interpreted as either a phenomenon of dedifferentiation or to microenvironmental selection of specific subclones. S-100 was equally expressed in the three contexts. The xenotransplant's ultrastructural features were those of a highly undifferentiated tumor. No significant immunophenotypic or ultrastructural differences between the two morphologically distinct populations were found. 4. Thus, our data demonstrate that NG97 cells constitute a pure glial-committed cell line, which may prove useful as a malignant glioma model in studies addressing pathophysiological, diagnostic and therapeutic issues.

Animals↗
PubMed2005

Developmental potential and ultrastructural injuries of metaphase II (MII) mouse oocytes after slow freezing or vitrification.

PURPOSE: The purpose of this study was to determine the developmental ability and ultrastructure of MII mouse oocytes after cryopreservation by slow freezing or vitrification. METHODS: Ovulated MII mouse oocytes were allocated to slow frozen, vitrified and control groups. Oocytes in the slow frozen and vitrified groups were cryopreserved using 1,2 propandiol (PROH) and ethylene glycol (EG) respectively as cryoprotectants. After thawing, the surviving MII oocytes in both cryopreserved groups and the control group were inseminated and their developmental ability was compared. The ultrastructure of MII oocytes in both cryopreserved groups was assessed immediately after thawing and 10 h post insemination at the pronuclear stage, and compared with that of the control group. RESULTS: The survival rates were nearly identical in both cryopreserved groups. The fertilization rates were also identical and comparable to that of the control group. The further development of vitrified oocytes was similar to that of the control oocytes, whereas it was severely limited in the slow-frozen oocytes. In the slow-frozen MII oocytes, the intermediate filaments were destroyed and the oolemma and microvilli were also modified. At the pronuclear stage deterioration of mitochondria and the presence of numerous vacuoles were also observed within the ooplasm. In the vitrified MII oocytes, the intermediate filaments were the only structures affected and these cytoskeletal elements were reorganized at the pronuclear stage. CONCLUSIONS: Vitrification results in less ultrastructural damage and better post fertilization development of MII mouse oocytes than slow freezing.

Animals↗