Immunomorphometric variations of sustentacular cells of the male viscacha adrenal medulla during the annual reproductive cycle. Effects of androgens and melatonin

The adrenal medulla is crucial for the survival of species facing significant environmental changes. The parenchyma is composed mainly of chromaffin cells, ganglion cells and sustentacular cells (SC). The male viscacha exhibits seasonal variations of gonadal activity and other metabolic functions. The aim of this work was to investigate the influence of the reproductive conditions on the morphology of SC of this rodent. In addition, the effects of testosterone and melatonin on these cells were studied. Immunoexpression of S100 protein, GFAP and vimentin were analyzed. Furthermore, the distribution of adrenergic and noradrenergic chromaffin cells subpopulations was studied for the first time in this species. SC present long cytoplasmic processes in contact with chromaffin cells, probably generating an intraglandular communication network. Significant differences (p?<?0.05) in the %IA (percentage of immunopositive area) for the S100 protein were observed according to winter (4.21?±?0.34) and summer (3.51?±?0.15) values. In castrated animals, the %IA (6.05?±?0.35) was significantly higher in relation to intact animals (3.95?±?0.40). In melatonin-treated animals the %IA (3.62?±?0.23) was significantly higher compared to control animals (2.65?±?0.26). GFAP immunoexpression was negative and no noradrenergic chromaffin cells were detected suggesting an adrenergic phenotype predominance. Vimentin was observed in SC, endothelial cells and connective tissue. Results indicate that SC exhibit variations along the annual reproductive cycle, along with castration and the melatonin administration. Our results suggest that in this rodent SC are not only support elements, but also participate in the modulation of the activity of the adrenal medulla; probably through paracrine effects.     

An immunocytochemical study of the development of the olfactory system in the three-spined stickleback (Gasterosteus aculeatus L., Teleostei).

Summary Antisera against a variety of substances have been found to produce an identical immunoreaction in the developing olfactory system of a teleost, the three-spined stickleback (Gasterosteus aculeatus). The label is localized in the olfactory placode, the olfactory nerve and those parts of the secondary olfactory tracts which constitute the dorsal descending fascicles and the ventral descending fibers of the medial olfactory tract. The label was first detected 3 days after fertilization (3D) in the olfactory placode where labeled supporting cells were observed. At 4D, the label was observed at the site of the developing olfactory bulbs. At 7D, the olfactory placode lost the direct contact with the brain and the labeled olfactory nerve became visible. At the same time, the medial olfactory tract emerged from the bulbs, and contacts with cells in the nucleus of the terminal nerve were observed. The development of the medial olfactory tract proceeded caudally, and by the end of I OD, the olfactory tract reached the periventricular hypothalamus. Pre-absorption of the antisera with the respective antigens did not abolish the capacity of the antisera to produce the label. The immunoreaction is thus not specific for the antigens against which the antisera have been raised. Yet the label produced by the immunoreaction is an extremely reliable marker for the primary olfactory tract, and the only existing marker by which secondary olfactory tracts can be visualized.     

Ultrastructural and morphometric study of the Sertoli cell of the viscacha (Lagostomus maximus maximus) during the annual reproductive cycle

Changes in the morphology of viscacha Sertoli cells were studied during the annual reproductive cycle. Sertoli cells exhibited marked nuclear and cytoplasmic changes. Seasonal variation in nuclear size and shape, chromatin texture, and nucleolus characteristics was observed. The seasonal patterns of the volume densities of the endoplasmic reticulum (ER), mitochondria, Golgi complex, dense bodies and lipid inclusions were distinct. Morphometric analysis revealed that the Golgi complex is the organelle most sensitive to seasonal change. It declined drastically in the regressed testes and its recovery was slow. The ER and mitochondria exhibited seasonal variations in their pattern and content, that was minimal during winter. In contrast, an accumulation of lipid and dense bodies, such as primary and secondary lysosomes, accompanied the spermatogenic arrest. The volume densities of both organelles were maximum during the restoration of spermatogenesis. The length and organization of the inter-Sertoli junctions also changed with the reproductive cycle. The Sertoli cell number per tubular cross section decreased significantly during the testicular regression, coincident with the presence of Sertoli cells with marked signs of involution. The degree of regression and recovery exhibited by the viscacha Sertoli cells was closely related to that shown by the associated germ cells. Therefore, seasonal endocrine fluctuations and local factors could be involved in the regulation of the morphological and functional characteristics of the viscacha Sertoli cells. These hormonal fluctuations are synchronized by the photoperiod through the pineal gland and its hormone, melatonin.     

Effects of an asymmetric triazine derivative,HOE 092 V,on Glugea anomala,Moniez, 1887 (Microsporidia) parasitic in the three-spined stickleback Gasterosteus aculeatus

 An asymmetric triazine derivative, HOE 092 V, 2-[3,5-α-dichloro-4-(4-methyl-sulfonylphenoxy)-phenyl]-1-methyl-hexahydro-1,2,4-triazine-3,5-dion, was tested in vivo against Glugea anomala parasitizing the connective tissue of sticklebacks (Gasterosteus aculeatus). Naturally infected sticklebacks were incubated in 10-l plastic aquaria in water containing 0, 2.5, 5, and 10 μg HOE 092 V/ml for 2, 3, and 4 h at 22°C. As seen at the ultrastructural level, the drug caused severe damage to all developmental stages of G. anomala except the mature spores. Starting with a dose of 2.5 μg/ml, the drug caused significant damage on uni- and multinucleate meronts, sporogonial plasmodia, and sporoblasts. The damage mainly consisted in a decrease in the number of ribosomes, an enlargement of the smooth endoplasmic reticulum and a vacuolization of the cytoplasma. When treatment was done with 5 μg for 2 h, multinucleate meronts and sporogonial plasmodia were no longer detectable, and the sporoblasts and the prespore stages except the mature spores had shrunk. After incubation of the infected fish with 10 μg HOE 092 V/ml and 4 h exposure, uninucleate meronts were no longer detectable by means of transmission electron microscopy. In the sporoblast mother cells, vacuolization of the cytoplasma and lysis of the nuclei occurred. However, mature spores were not affected. It seems likely that HOE 092 V can be successfully applied in medicinal baths against Microsporidia in fish. The infected fish should be incubated in separate, aerated containers. Received: 15 July 1995 / Accepted: 15 August 1995     

Ultrastructural and histochemical characterization of marmoset (Callithrix jacchus) Leydig cells during postnatal development

Summary Leydig cell development was investigated in the marmoset (Callithrix jacchus) testis from 24 h post partum (pp) up to sexual maturity, using ultrastructural and histochemical methods.Electron microscopically three different Leydig cell (LC) types could be distinguished: neonatal, immature and adult LCs. Neonatal LCs exhibited a round nucleus, large tubular mitochondria, abundant smooth endoplasmic reticulum (SER), and few small fat vesicles. The typical immature LCs showed a lobulated nucleus, smaller tubulo-vesicular mitochondria, less SER, and larger fat vesicles. Adult LCs contained an invaginated nucleus, tubular mitochondria, abundant SER, large fat vesicles, and lipofuscin granules. Neonatal Leydig cells occurred from 24 h pp up to 10 weeks pp, immature LCs from 2 weeks pp up to 20 weeks pp, and adult LCs from 20 weeks onwards. The appearance of a new LC population was accompanied by a numeric decrease and occurrence of regressive cells of the previous population. Developmental steps of differentiation were recognizable in the population of immature and adult LCs by the amout of SER (immature LCs), the size of mitochondria (adult LCs), and the morphology of the nucleus at the beginning of their appearance. Activity of 3-hydroxysteroid dehydrogenase (3-HSD) was moderate in immature LCs from 2 weeks pp to 12 weeks pp and strong in adult LCs. Neonatal LCs were not stained nor were immature LCs from 16 weeks to 20 weeks. Comparison of the ultrastructural classification of LCs and the activity of 3-HSD showed that the capacity for steroid biosynthesis does not necessarily include morphological differences. Activity of non-specific esterase (nE) was found in LCs except of the period between 16 and 25 weeks pp. A transient decrease of activity was seen during the first week of life. Both, the phase of decrease and the phase of inactivity were followed by distinct activation of nE. Lipid content and the size of single fat droplets in LCs, as seen with Sudan black staining, varied during postnatal development. Finely stained granules were typical for neonatal LCs, whereas larger droplets were found in immature and adult cells. Particularly high amounts of lipids were seen at one week and between 32–60 weeks pp, which was the regression of neonatal LCs, showing ultrastructurally higher amounts of lipids than intact neonatal LCs. High amounts of lipid during puberty (32–60 weeks pp) accompanied by an activation of nE at the onset of puberty suggest an initiative role in steroidogenesis. Based upon histochemical data four phases of LC development can be distinguished: a neonatal phase (24 h-7 days pp), and infantile phase (2–12 weeks pp), a phase of inactivity (16–25 weeks pp) and a puberral/adult phase (32 weeks pp-adult). In view of different morphological features, of degenerative LCs, of the simultaneous occurrence of neonatal and immature or immature and adult LCs for an extended period of time, and of histochemical results, it appears likely that all three Leydig cell populations differentiate from mesenchymal intertubular cells, but not from the previous population.This investigation was supported by the Deutsche Forschungsgemeinschaft (Sfb 174)     

A quantitative electron microscopic study of the effect of glucocorticoids in vivo on the early postnatal differentiation of paraneuronal cells in the carotid body and the adrenal medulla of the rat

Summary The postnatal differentiation of carotid body chief cells and endocrine adrenal medullary cells was comparatively examined during ontogenesis and in rats which were treated with dexamethasone for 7 days after birth. Ultrastructure and innervation of carotid body chief cells are mature in neonates according to the functional requirements of chemoreception. By the end of the first postnatal week, only an increase in number of dense core vesicles can be noticed, the concentration of which then will reach the adult level. Under the effect of dexamethasone most of the heterochromatin is transformed into finely dispersed euchromatin within the nuclei of carotid body chief cells. In the cytoplasm, the Golgi apparatus becomes larger and the granular endoplasmic reticulum hypertrophic. The number of catecholamines storing dense core vesicles increases considerably. The innervation density remains constant. In contrast to the carotid body chief cells, the adrenal medullary cells have not reached their definitive maturity at the time of birth. Besides phenotypes of adrenaline-cells, noradrenaline-cells and small granules containing cells, pheochromoblasts and intermediary cells can be seen as well. Their cytoplasm is sparse, the concentration of dense core vesicles and the innervation density very low. After 8 days of postnatal ontogenesis, pheochromoblasts and intermediary cells are no longer present in the adrenal medulla. In adrenaline-cells and noradrenaline-cells, important processes of growth can be noticed, the cytoplasm has grown in extent, the number of dense core vesicles doubled and the innervation density of single cells triplicated. Only the few small granules containing cells remain small. Under the effect of dexamethasone also in the nuclei of chromaffin cells a transformation of heterochromatin into euchromatin occurs. The increase in number of dense core vesicles is relatively lower than in carotid body chief cells. The significant growth of innervation density during the first postnatal week was inhibited. Our observations suggest that dexamethasone stimulates the synthesis of catecholamines in adrenal medullary cells of newborn rats less pronouncedly than in carotid body chief cells. This could be attributed to the inhibited formation of synapses of growing chromaffin cells and to the in vivo active endocrine counter-regulation.     

Ultrastructural study of benign, low-malignant potential (LMP), and malignant ovarian tumors

Ultrastructural characteristics of benign, low-malignant potential (LMP), and malignant ovarian tumors were investigated, considering the aspects of histologic subtypes and histologic grading. In addition, the histogenesis of ovarian cancer was histologically investigated in an attempt to elucidate whether malignant tumor was generated from benign or LMP tumor, or whether it was generated de novo from normal tissues. Although all the benign, LMP, and malignant tumors appeared to be derived from Mullerian duct in serous tumors, the origin of endometrioid or mucinous tumor could not be ultrastructurally clarified. However, there was ultrastructural similarity between benign and malignant tumors among serous, endometrioid, and mucinous tumors, and it was suggested that benign adenoma may be the developmental origin of malignant tumors regardless of the histologic subtype. In addition, the investigation of endometrioid tumors revealed that the differences of histologic grading in malignant tumors reflected the ultrastructural differences, and that G1 tumor had an ultrastructure that was more similar to that of benign and LMP tumors than to that of G2 tumor.     

Ultrastructural study of Sertoli cells in rat seminiferous tubules during intrauterine life and the postnatal period

Summary Sertoli cells have various functions: mechanical (creation of two compartments in the seminiferous tubules, migration of germinal cells), secretory (secretion of anti-Müllerian hormone, inhibin, androgen-binding-protein and estrogen) and phagocytic.We report an ultrastructural study of the rat Sertoli cell during maturation and consider possible correlations between the acquisition of certain morphological characteristics and certain functions.During fetal life, the Sertoli cell possesses differentiated zones of junction with the gonocytes and seems to have a role in the migration of the gonocytes towards the periphery of the seminiferous tubule. The Sertoli cell performs the phagocytosis of the gonocytes which degenerate during their migration, and seems to be the site of production of protein granules, whose presence can be related to the synthesis of anti-Müllerian hormone.After birth and before puberty, when the inclusions resembling secretory granules disappear, the Sertoli cell membranes in contact with spermatocytes II and spermatids differentiate, forming, through the differentiated junctional complexes, two compartments (adluminal and luminal) in the seminiferous tubules. Finally, they acquire the characteristics of active secretory cells, capable, in particular, of steroid synthesis.     

A novel subtyping of intestinal metaplasia of the stomach,with special reference to the histochemical characterizations of endocrine cells

Summary Intestinal metaplasia of the stomach was grouped into 3 subtypes (A, B and C) according to the degree of pyloric gland involution which was judged from patterns of paradoxical Concanavalin A staining after Katsuyama and Spicer. The appearance of endocrine cells was investigated with immunohistochemical and silver methods. Type A metaplasia with slightly to moderately atrophic pyloric glands corresponded to the incomplete type in the previous classification, while Type C showing complete disappearance of pyloric glands corresponded to the complete type. Type B with severely atrophic pyloric glands was an intermediate. This subtyping reflects the cell kinetics in the intestinalized mucosa well. Regarding the endocrine cells, their total number varied in the order Type A > Type B > Type C. The selective populations of the endocrine cells including glicentin-containing cells, Grimelius-positive argyrophil cells without argentaffinity and intestinal-type enterochromaffin cells frequently formed hyperplastic foci in the intestinalized areas, where the other gut-type and proper gastric-type endocrine cells were scarcely noted. Immunoreactivity of glucagon or bovine pancreatic polypeptide were occasionally identified in a subpopulation of the glicentin-containing cells.     

大鼠老年多器官功能障碍综合征生理、生化及超微结构的变化

目的:研究老年多器官功能障碍综合征(MODSE)动物模型超微结构的改变特点与生理、生化指标变化,探讨老年多器官功能障碍发生机制,提供防治老年多器官功能障碍综合征的依据。方法:建立盲肠结扎穿孔术(CLP)的MODSE组、青年多器官功能障碍综合征(MODSY)组及相应的假手术对照组。在术后24h采用透射电镜技术观察重要器官(心、脑、肺、肝和肾)的超微结构变化,同时检测各组大鼠的呼吸频率(RR)、心率(HR)和血压(BP)、血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、总胆红素(TB)、肌酐(Cr)、尿素氮(BUN)等生理、生化指标水平的改变。结果:观察MODSE组和MODSY组的重要器官组织超微结构时发现,术后24h两组实验大鼠的重要器官及其血管内皮细胞的超微结构均有不同程度的损伤;除RR、BP、HR指标外,其他5项生化指标较假手术对照组亦有显著差异(P＜0.05)。结论:不同程度的重要器官组织的细胞损伤(其中以MODSE实验组的细胞损伤最为严重)与相应的生理、生化指标改变的一致性,说明血管内皮细胞的损伤可不同程度导致微循环障碍,引起相应组织的供血不足,进而使细胞的代谢功能受到影响,发生细胞变性、坏死。在纠正器官功能不全的同时,尤应注意积极采取保护血管内皮细胞的治疗措施。     

Redistribution of membranes and cytoskeletal proteins in chicken oxyntic cells during the HCl secretory cycle: Ultrastructural and immunofluorescence study

Changes in ultrastructure and cytoskeletal organization by avian oxyntic cells, at the onset of HCl secretion, were analysed. Cells in resting state, induced by fasting and cimetidine, were compared with histamine stimulated secreting cells. Ultrastructural studies were done by transmission electron microscopy; the distribution of prekeratin, myosin, and filamin-like protein, by immunofluorescence; and that of F-actin using FITC-phalloidin. Resting cells show short pericellular clefts. These are increasingly deepened in secreting cells by a reorganization of the lateral cell borders involving displacement of the junctional complexes toward the cell base and incorporation of the tubular system to the luminal plasma membrane. In secreting cells, the processes of the secretory surface are concentrated in a pericellular groove. Histamine stimulation induces a drastic redistribution of cytoskeletal proteins. In chicken oxyntic cells, in addition to the F-actin cytoskeleton associated with the membranes of the secretory surface, there is a cytoskeletal ring containing F-actin, myosin, and a filamin-like protein, located at the level of the junctional complexes. In resting cells, filaments and masses of cytoskeletal matrix are associated with the zonula adherens. In secreting cells, the junctional complexes maintain their association with the filamentous ring, while the amorphous matrix is replaced by microfilaments that support the processes of the luminal surface. Intermediate filaments form a peripheral ring probably associated with the zonula adherens, and project from the ring toward the cell cytoplasm. Thus, with the onset of HCl secretion, the apical cytoskeletal ring of resting cells displaces toward the cell base. A role for this cytoskeletal ring in the changes in shape parallel to HCl secretion is discussed.     

Ultrastructural study on the follicle-associated epithelium of nasal-associated lymphoid tissue in specific pathogen-free (SPF) and conventional environment-adapted (SPF-CV) rats

Membranous (M) cells in follicle-associated epithelium (FAE) play an important role in the mucosal immunity through transport of a variety of foreign antigens to the underlying mucosa-associated lymphoid tissue (MALT). We aimed to investigate the ultrastructure of M cells in the FAE covering nasal-associated lymphoid tissue (NALT) both in specific pathogen-free (SPF) rats and in conventional environment-adapted (SPF-CV) rats aged 8–38 wk. In NALT of both SPF and SPF-CV rats, FAE included the nonciliated microvillous cell, which appears to be an analogue of M cell previously described in other MALT. In SPF rats, M cells increased in number only slightly with age, and they maintained morphological uniformity irrespective of age. In SPF-CV rats, M cells selectively increased in number resulting in prominent expansion of FAE surface area in parallel with the duration of maintenance in a conventional environment. In addition, M cells in SPF-CV rats showed heterogeneity in their surface morphology such as the length and number of microvilli and cell surface area and outline. In addition, the FAE was stratified by various subtypes of M cells, which were characterised by several subcellular alterations including the presence of many keratin filaments, homogeneous dark bodies and extensive cytoplasmic interfoliation with wide intercellular spaces filled with amorphous proteinaceous material. These characteristics of M cells in SPF-CV rat were intimately related with a preferential influx of immunocompetent cells into the FAE, which was not seen or was very rare in SPF rats irrespective of age. The results suggest the possibility that NALT may effectively carry out the mucosal immune response against antigenic stimuli of different magnitude through the unique dynamics of M cells which seem to be influenced by the infiltration of immunocompetent cells.     

A carbohydrate histochemical study on surface mucous cells, mucous neck cells and chief cells in the gastric mucosa of developing mice

The ontogenesis of the mouse gastric mucosa was studied by carbohydrate histochemistry and 3H-thymidine autoradiography. Surface mucous cells and glandular cells were identified from day 16 of gestation. Sugar residues in the mucin of surface mucous cells seem to undergo no major changes throughout the period under study, since secretory granules of the cells were positive in periodic acid-Schiff (PAS) and galactose oxidase-Schiff (GOS) reactions and consistently bound certain lectins. Chief cells and mucous neck cells are not separated until the third postnatal week, though primitive chief cells are present during earlier developmental stages. Secretory granules of primitive chief cells shared positive PAS and GOS reactions with mucous neck cells and bound similar lectins, but the intensity was generally weaker. The granules of primitive chief cells were also stained by Bowie's solution which exclusively stained zymogen granules in chief cells in adults. These results suggest that secretory granules of primitive chief cells contain a complex carbohydrate similar to mucin in mucous neck cells, but with a lower sugar/protein ratio. It is concluded by studies using 3H-thymidine autoradiography combined with carbohydrate histochemistry that, though immature surface mucous cells, primitive chief cells and mucous neck cells actively proliferate, chief cells rarely undergo mitosis.     

骨髓基质干细胞向神经细胞诱导分化及其机制的研究进展

骨髓基质干细胞是一种存在于骨髓间质中具备多向分化潜能的成体干细胞,其不仅能分化为中胚层起源的骨、软骨和脂肪细胞,还可以在特定条件下诱导分化为神经外胚层起源的神经细胞。主要对骨髓基质干细胞向神经细胞诱导分化及其机制的研究进展作一综述。     

Differential localization and expression of urokinase plasminogen activator (uPA), its receptor (uPAR), and its inhibitor (PAI-1) mRNA and protein in endometrial tissue during the menstrual cycle

Normal endometrium is a highly dynamic tissue, which responds to ovarian steroids with cyclic proliferation, differentiation (secretion), and degradation (menstruation). The urokinase plasminogen activator (uPA)-dependent proteolytic cascade as well as ligand activation of the uPA receptor (uPAR) is critically involved in physiological as well as pathophysiological aspects of tissue expansion and remodelling. Cyclic variation and distribution of uPA, uPAR and plasminogen activator inhibitor 1 (PAI-1) mRNA were examined by in situ hybridization, real-time PCR and northern blot in normal endometrium. Their corresponding proteins were localized with immunohistochemistry. uPA mRNA is exclusively expressed by stromal cells, whereas uPA protein is present in both epithelial and stromal cells. Immunostaining for uPA protein is reduced or undetectable at midcycle, thus coinciding with peak concentration of uPA in the uterine fluid. uPAR mRNA is expressed by epithelial cells in the proliferative phase and by stromal cells in the secretory phase. However, epithelial cells stain for uPAR protein throughout the cycle, suggesting that uPAR may detach from stromal cells and then bind to epithelial cells in the secretory phase. PAI-1 mRNA is located in vessel walls. The late secretory phase has greatly increased expression of all three mRNA and their proteins, mainly in pre-decidual cells in the superficial stroma. Discordant localization of the mRNA and proteins suggest that uPA is produced by stromal cells, released and bound to epithelial cells in both the proliferative and secretory phases, whereas uPAR is released from the stroma and bound to epithelial cells in the secretory phase. Also, the present data together with earlier reports suggest that uPA is released from the epithelial cells to the uterine fluid.     

An immunocytochemical study of the endocrine cells in the stomach and duodenum of Zonotrichia capensis subtorquata (Passeriformes, Emberizidae)

The main purpose of this study was to evaluate the regional distribution pattern and relative frequency of some endocrine cells in the three portions of the gastrointestinal tract (GIT)--the proventriculus, gizzard and duodenum- of the rufous-collared sparrow (Zonotrichia capensis subtorquata), by immunohistochemical methods using six types of polyclonal antisera, specific for serotonin (5-HT), somatostatin (D cells), glucagon, motilin, polypeptide YY (PYY) and insulin. In the proventriculus, endocrine cells immunoreactive for all of these markers were observed. The somatostatin-immunoreactive cells were found with greater frequency, with the presence of cytoplasmic processes. In the gizzard, endocrine cells secreting somatostatin, 5-HT and PYY were detected, while those secreting glucagon and insulin were not. In the final part of the gizzard, endocrine cells secreting 5-HT were more frequent, and cells secreting somatostatin and insulin were not detected. All of the cell types studied were observed in the duodenum in different frequencies, except for cells immunoreactive for glucagon and insulin. The somatostatin-positive (D cells) were the most numerous, being more prevalent in the intestinal glands. The other endocrine cells were identified in smaller numbers, some of them located in the intestinal villi and Lieberkuhn glands. The finding of these cell types in the duodenum confirms their preferential location in the final portions of the principal segments of the digestive system and suggests control by feedback of its functions. In conclusion, some interesting distributional patterns of gastrointestinal endocrine cells were found in this species of sparrow.     

MCMV影响神经干细胞cyclins表达和细胞周期进程的体外实验研究

目的:观察鼠巨细胞病毒(MCMV)感染对体外培养神经干细胞(NSCs)细胞周期进程和cyclins表达的影响,探讨MCMV感染致脑发育异常的机制。方法:本实验体外分离、培养和鉴定BABL/C胎鼠NSCs,以感染复数(MOI)为5,1和0.1的MCMVsmith毒株感染NSCs并于感染后1,2,3,4,5,6d收集细胞,采用Cyclin/DNA双参数流式细胞术检测感染细胞cyclinA,cyclinB1,cyclinD1,cyclinE的表达和细胞周期时相的动态变化,观察MC-MV对感染NSCs细胞周期进程的影响。结果:体外分离培养的NSCs呈球样生长,神经干细胞特异性标记Nestin表达阳性,并可进一步分化为GFAP阳性的星形胶质细胞和NF-200阳性的神经元;各感染组cyclinA,cyclinB1,cy-clinD1和cyclinE的表达均上调,其中MOI=0.1表达逐渐上升,在第6d达峰值,MOI=1组表达高峰在第4d,MOI=5组表达高峰在第3d;感染组G0/G1期细胞比率减少,S期和G2/M期细胞比率增加,其变化趋势与cyclins的表达基本一致,并随MOI的增加变化越明显。结论:Cyclin/DNA多参数流式细胞术可用于NSCs细胞周期的分析;MCMV可通过上调cyclinA,cyclinB1,cyclinD1,cyclinE的表达来影响NSCs细胞周期进程,并与MOI存在一定量效依赖关系;MCMV感染可诱导NSCs从G0/G1期进入S期,出现S期和G2/M期偏移和阻滞,影响NSCs细胞周期进程,这可能是CMV抑制NSCs增殖并导致先天性脑发育异常的重要机制之一。     

Evaluation of PvuII and XbaI polymorphisms in the estrogen receptor alpha gene (ESR1) in relation to menstrual cycle timing and reproductive parameters in post-menopausal women

Objective: To evaluate the association of −397T>C and −351A>G single nucleotide polymorphisms (SNPs) – also called PvuII and XbaI, respectively – located on estrogen receptor alpha (ERS1) gene with age at menarche, menopause onset, fertility and miscarriage in a population of post-menopausal women. Study design: Cross-sectional study with 273 healthy, high miscegenated, post-menopausal women (mean age of 63.1 ± 9.7 years old). Subjects were genotyped for PvuII and XbaI SNPs by PCR-RFLP and confirmed by automatic sequencing. Reproduction informations (age at menarche, age at menopause, number of pregnancies, fertility rate and miscarriages) were obtained by retrospective study using a questionnaire. Result(s): Age at menarche, menopause onset, number of pregnancies, total fertility rate, and parity did not seem to be influenced by any of the studied genotypes (chi-square, p > 0.05). However, women carrying the xx genotype showed a 44% higher chance of miscarriage, whereas this value did not trespass 16% for any other genotype analyzed. It has been also observed a higher occurrence of miscarriage in association with combined xxpp genotype of ERS1 gene (chi-square, p < 0.01). Conclusion(s): The present data indicate that the studied SNPs on ERS1 gene do not influence the menstrual cycle timing and parity but there is a strong relationship between the xx ERS1 SNP genotype and the incidence of miscarriage in the post-menopausal population analyzed.