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The sexually dimorphic larynx of Xenopus laevis: development and androgen regulation. 非洲爪蟾两性二型喉:发育和雄激素调节。
Pub Date : 1986-12-01 DOI: 10.1002/aja.1001770404
D Sassoon, D B Kelley

The aims of this study were to characterize sexual dimorphism in the larynx of adult Xenopus laevis and to determine how sex differences arise during postmetamorphic development. The larger male larynx is a result of greater cell numbers in both cartilage and muscle. The dilator laryngis muscle of the male larynx has 6-7 times more muscle fibers than that of the female. At metamorphosis, the larynx is sexually monomorphic and feminine in phenotype. The DNA content of the male larynx doubles during the first 6 months following metamorphosis; there is no net DNA increase in the female larynx during this time. Both sexes experience a marked increase in laryngeal DNA content and mass between 6 months and adulthood. The number of muscle fibers in the male larynx increases at an average rate of 150 fibers a day during the first 10 months of postmetamorphic development. There is no net change in fiber numbers in the female larynx from metamorphosis to adulthood. Administration of the antiandrogen Flutamide to metamorphic frogs prevents the net addition of laryngeal muscle fibers in males. Thus, we propose that addition of postmetamorphic laryngeal muscle fibers in males is dependent upon the presence of circulating androgens. Exogenous testosterone administration results in an increase in laryngeal mass, DNA content, and cellular proliferation in juvenile frogs. Using [3H]thymidine injections to probe ongoing, as well as testosterone-induced, cell proliferation, we conclude that cellular proliferation is regulated differently in males and females during development. Thus androgen-induced proliferation is one cellular mechanism responsible for the sexual dimorphism observed in adults.

本研究的目的是表征成年非洲爪蟾喉部的性别二态性,并确定性别差异是如何在变态发育过程中产生的。男性喉部较大是由于软骨和肌肉中细胞数量较多。男性喉部扩张肌的肌纤维是女性的6-7倍。在变态时,喉是两性单形的,在表型上是女性的。雄性喉部DNA含量在变态后的前6个月翻倍;在此期间,女性喉部没有净DNA增加。在6个月到成年期间,男女喉部DNA含量和质量都显著增加。在变态发育后的头10个月,男性喉部肌肉纤维的数量以平均每天150根纤维的速度增加。从变态到成年,女性喉部的纤维数量没有净变化。给变性蛙服用抗雄激素氟他胺可防止雄性喉肌纤维的净增加。因此,我们提出,男性变态后喉肌纤维的增加取决于循环雄激素的存在。外源性睾酮管理导致喉肿块,DNA含量的增加,并在幼年青蛙细胞增殖。使用[3H]胸腺嘧啶注射来探测正在进行的细胞增殖,以及睾酮诱导的细胞增殖,我们得出结论,在发育过程中,男性和女性的细胞增殖受到不同的调节。因此,雄激素诱导的增殖是导致在成人中观察到的两性异形的一种细胞机制。
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引用次数: 115
Somitic-vertebral correlation and vertebral levels in the human embryo. 人胚胎中躯体-椎体的相关性和椎体水平。
Pub Date : 1986-09-01 DOI: 10.1002/aja.1001770103
F Müller, R O'Rahilly

Somitic and vertebral interrelationships and levels were studied in 84 human embryos of stages 9-23 (3-8 postovulatory weeks). The first four somites are occipital, the occipitocervical junction is at somites 4/5, and eight somites are involved in the cervical region: X, Y, Z, and C. 3-7. By stage 17 the total number of occipitovertebral "units," namely 38 or 39, is attained. Resegmentation (Neugliederung) of sclerotomes is not supported. A new scheme of somitic/vertebral correlation is proposed in which somites and centra are in register. Differential growth of the regions of the vertebral column was calculated, and it was found that the percentages of the total column occupied by the various regions vary from one stage to another. The cervical and coccygeal regions decrease, the thoracic and lumbar regions increase, and the sacral region remains more or less constant during embryonic development. The following structures descend with reference to the vertebral column during the embryonic period proper: roots of lower limbs, thyroid gland and thymus, tracheal bifurcation, lungs, heart, diaphragm, abdominal arteries, mesonephroi, and suprarenal glands. The gonads may descend slightly. The scapulae and the separation point between the trachea and the esophagus remain at a fairly constant level. The metanephroi ascend. The migration of many of these structures (e.g., the heart, diaphragm, and metanephroi) is much more marked in the embryonic period than later although it continues during the fetal and postnatal periods. The conus medullaris ascends during the fetal period. Anomalies of migration that affect such organs as the thyroid gland, gonads, and metanephroi are discussed.

研究了84个9-23期(排卵后3-8周)人类胚胎的体和椎体相互关系和水平。头4个小体是枕部,枕颈连接处在4/5小体,颈区有8个小体:X、Y、Z、c - 3-7。到第17阶段,枕椎“单元”的总数达到38或39。不支持硬切块的再分割。提出了一种新的体/椎体相关方案,其中体和椎体在寄存器中。计算了脊柱各区域的差异生长,发现各区域占脊柱总生长的百分比在不同阶段有所不同。在胚胎发育过程中,颈椎和尾骨区域减少,胸椎和腰椎区域增加,骶骨区域或多或少保持不变。在胚胎期,以下结构相对于脊柱下降:下肢根部、甲状腺和胸腺、气管分叉、肺、心脏、隔膜、腹动脉、肾中膜和肾上腺。性腺可能会轻微下降。肩胛骨和气管和食道之间的分离点保持在一个相当恒定的水平。后肾上升。许多这些结构(如心脏、横膈膜和后肾)的迁移在胚胎期比后期更为明显,尽管它在胎儿期和出生后仍在继续。在胎儿时期,髓圆锥上升。对影响甲状腺、性腺和后肾等器官的异常迁移进行了讨论。
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引用次数: 49
The renal cortical lymphatic system in the rat, hamster, and rabbit. 大鼠、仓鼠和家兔的肾皮质淋巴系统。
Pub Date : 1986-09-01 DOI: 10.1002/aja.1001770104
G K Niiro, H M Jarosz, P J O'Morchoe, C C O'Morchoe

Rat, hamster, and rabbit renal cortical lymphatics were examined by light and electron microscopy. Rat and hamster kidneys possessed both intra- and interlobular lymphatics that were structurally similar at the light microscopic level. Ultrastructural examination of the hamster lymphatic endothelium, however, revealed an unusual arrangement of cytoplasmic extensions not seen in the other two species. The intralobular lymphatics were related primarily to tubules, afferent arterioles, and renal corpuscles and were consistent with lymph formation from both plasma filtrate and tubular reabsorbate. Interlobular lymphatics were seen in connective tissue associated with the interlobular blood vessels. Rabbit cortex contained only interlobular lymphatics. Cross-sectional area, maximum diameter, volume density, and profile density were determined by stereological measurements using a computer-based image analyzer. The morphological data from the rat were used, in combination with published values for lymph flow, to calculate the rate of lymph formation per unit area of endothelium in lymphatics of the renal cortex. Among kidneys fixed by retrograde perfusion, the cortical lymphatic system was most extensive in maximum diameter, volume density, and profile density. It was smallest in the rabbit and intermediate in the rat. Lower volume and profile density were found for rat kidneys fixed by the dripping technique. It was concluded that: tubular reabsorbate probably contributes to renal lymph in the rat and hamster, but not in the rabbit; significant differences exist in the extent of the renal lymphatic systems among the three species, with the hamster kidney having the richest network and the rabbit the poorest; the method of fixation influences the measured size and density of renal cortical lymphatics; and the estimated rate of lymph formation in the kidney of the rat is roughly comparable to that in the dog.

用光镜和电镜检查大鼠、仓鼠和家兔肾皮质淋巴管。大鼠和仓鼠的肾脏都具有小叶内和小叶间淋巴管,在光镜下结构相似。然而,对仓鼠淋巴内皮的超微结构检查显示了一种不寻常的细胞质延伸排列,在其他两个物种中未见。小叶内淋巴管主要与小管、传入小动脉和肾小体有关,与血浆滤液和小管再吸收液形成的淋巴管一致。小叶间淋巴管见于与小叶间血管相关的结缔组织。兔皮质仅含小叶间淋巴管。横截面积、最大直径、体积密度和剖面密度由基于计算机的图像分析仪的立体测量确定。利用大鼠的形态学数据,结合已公布的淋巴流量值,计算肾皮质淋巴管每单位面积内皮细胞的淋巴形成率。在逆行灌注固定的肾脏中,皮质淋巴系统在最大直径、体积密度和剖面密度上最为广泛。兔体内最小,大鼠体内居中。采用滴灌法固定大鼠肾脏,体积和密度较低。结论:肾小管重吸收可能对大鼠和仓鼠肾淋巴有贡献,对家兔无贡献;三种动物肾淋巴系统的范围存在显著差异,仓鼠肾淋巴网络最丰富,家兔肾淋巴网络最贫乏;固定方法影响肾皮质淋巴管的大小和密度;估计大鼠肾脏中淋巴形成的速率与狗的大致相当。
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引用次数: 6
In vivo exocytosis of lysosomes by the endothelium of the venous sinuses of bone marrow and liver: visualization at normal and low body temperature. 骨髓和肝脏静脉窦内皮在体内溶酶体的胞吐:在正常和低体温下的可视化。
Pub Date : 1986-09-01 DOI: 10.1002/aja.1001770105
P P De Bruyn, Y Cho

We have visualized the exocytosis of lysosomes into the peripheral circulation by the phagocytic endothelia of the venous sinuses of liver and bone marrow of rats. Perfusion fixation at normal body temperature produced images of the earliest stages of lysosomal exocytosis. After fixation at low body temperatures (7-12 degrees C), advanced stages of this process became evident, showing extrusion of lysosomes and their contents into the circulation. It is postulated that this form of exocytosis has escaped structural detection because of its rapidity and relative infrequency as compared to merocrine secretory exocytosis, and that fixation at low body temperatures arrests or slows down these exocytic events in sufficient measure for ultrastructural visualization. The possibility that this lysosomal exocytosis contributes to the presence of lysosomal enzymes detected in the peripheral blood should be considered. In addition, it is likely that lysosomal degradation products may be discharged by exocytosis into the circulation.

我们观察了溶酶体通过大鼠肝脏和骨髓静脉窦的吞噬内皮向外周循环的胞吐。在正常体温下灌注固定产生溶酶体胞吐最早阶段的图像。在低体温(7-12摄氏度)下固定后,这个过程的高级阶段变得明显,显示溶酶体及其内容物进入循环。据推测,与分泌性分泌胞吐相比,这种形式的胞吐因其快速和相对较少而未被结构检测到,并且在低体温下固定足以阻止或减缓这些胞吐事件,以实现超微结构可视化。这种溶酶体胞吐作用有助于外周血中检测到溶酶体酶的存在的可能性应予以考虑。此外,溶酶体降解产物可能通过胞吐作用排入血液循环。
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引用次数: 9
Mesenchyme formation from the trigeminal placodes of the mouse embryo. 小鼠胚胎三叉神经基间质形成。
Pub Date : 1986-05-01 DOI: 10.1002/aja.1001760103
D H Nichols

The trigeminal placode is a thickened region of ectodermal epithelium located along the side of the embryonic head. Mesenchyme escapes from the placode to form neurons of the trigeminal (V) ganglion. To further our knowledge of the morphogenesis of this escape, plastic thick sections were cut from mouse embryos and stained for light microscopy by using a technique which revealed escaping mesenchyme. The escape of trigeminal mesenchyme began at approximately 12 somites of age and was substantially complete by 30 somites. These results provided spatial/temporal orientation for a subsequent electron microscopic study. The first ultrastructural manifestation of escape was the penetration of an otherwise continuous basal lamina by small cell processes. The presence of longitudinally oriented microtubules within these processes suggests that mesenchymal cells escape through the basal lamina by using microtubules to direct/move their contents (e.g., the cell nucleus) into an enlarging process. Nuclei were distorted as they passed into these processes. This distortion suggests that basal lamina, together with a possible contribution from basal microfilaments, forms a rigid obstruction which is disrupted in the region from which a process is formed. In some cases a collar of basal lamina was observed around the necks of processes, but their distal membranes were invariably lamina-free. This lamina-free membrane is possibly that which is newly formed to accommodate the growing process. In later stages of escape, instances were observed in which the lamina was completely absent beneath an escaping cell and partially degraded beneath adjacent cells as well. These instances suggest that enzymatic digestion may play a role in degrading the lamina during mesenchymal escape. Apical desmosomes were often retained beyond the initial stages of escape. Mechanisms involved in their disruption are thus not among those which initiate escape.

三叉基是位于胚胎头侧的外胚层上皮增厚区。间质从基板逃逸形成三叉神经节的神经元。为了进一步了解这种逃逸的形态发生,我们从小鼠胚胎上切下塑料厚切片,并在光镜下使用一种显示逃逸间质的技术进行染色。三叉神经间质在大约12岁时开始逃逸,到30岁时基本完成。这些结果为随后的电子显微镜研究提供了空间/时间方向。逃逸的第一个超微结构表现是小细胞突穿透连续的基板。纵向定向微管在这些过程中的存在表明,间充质细胞通过微管引导/移动其内容物(如细胞核)进入增大过程,从而通过基底层逃逸。原子核在进入这些过程时是扭曲的。这种扭曲表明,基底层连同可能来自基底层微丝的作用,形成了一种刚性障碍,这种障碍在形成过程的区域被破坏。在某些情况下,在突颈周围观察到基底膜的项圈,但它们的远端膜总是无层。这种无层膜可能是为了适应生长过程而新形成的膜。在逃逸的后期,观察到在逃逸细胞下的层完全缺失,在相邻细胞下的层也部分降解的情况。这些实例表明,酶消化可能在间充质逃逸过程中对膜的降解起作用。顶端桥粒通常在脱出的初始阶段后仍保留。因此,破坏它们的机制并不在引发逃逸的机制之列。
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引用次数: 42
Morphometric analysis of circadian variations in the retinal photoreceptor synaptic terminals of the adult and fetal guinea pig. 成年和胎儿豚鼠视网膜光感受器突触末端昼夜节律变化的形态计量学分析。
Pub Date : 1986-05-01 DOI: 10.1002/aja.1001760102
M D McCartney, D H Dickson

In order to test whether the alterations in photoreceptor synaptic terminal size and shape reported in lower vertebrates occur in a mammalian visual system, adult and fetal guinea pig retinas were exposed to an LD 12:12 lighting cycle, as well as to long-term light (LL) and long-term dark (DD) regimes. Representative random samples from all retinal quadrants, obtained at various times during these lighting regimes, were processed for electron microscopy. The synaptic terminals of all three photoreceptor cell types in this retina (alpha and paranuclear rods, and cones) were analyzed with computer-assisted morphometrics for changes in their area, perimeter, synaptic vesicle density, and the degree of plasmalemmal infolding. The data showed all three types of adult receptor terminals to have increased area and vesicle density, as well as decreased membrane infolding, during the light period, while both types of rods showed increased perimeter measurements in the dark. Results from adults maintained under extended lighting conditions (LL and DD) showed no difference when compared with sample times during a typical LD 12:12 lighting regimen where clear statistical differences existed. Data from fetal retinas showed no significant sustainable pattern in any of the measured variables. These quantitative findings have led to the conclusion that while alterations in perimeter measurements may be explained by using the vesicle recycling hypothesis, observed changes in terminal size and shape may be controlled by a light-initiated or light-enhanced mechanism and effected through an annular configuration of cross-striated fibrils found within these photoreceptor synaptic terminals.

为了测试在低等脊椎动物中报道的光感受器突触末端大小和形状的改变是否发生在哺乳动物的视觉系统中,将成年和胎儿豚鼠的视网膜暴露在LD 12:12的光照周期中,以及长期光照(LL)和长期黑暗(DD)的环境中。在这些光照条件下的不同时间获得的所有视网膜象限的代表性随机样本进行了电子显微镜处理。用计算机辅助形态测量学分析视网膜中所有三种感光细胞类型(α和副核杆细胞和视锥细胞)的突触末端的面积、周长、突触囊泡密度和浆层内折叠程度的变化。数据显示,在光照期,所有三种类型的成年受体末端的面积和囊泡密度都增加了,膜折叠也减少了,而在黑暗期,两种类型的杆状体的周长都增加了。与典型的LD 12:12照明方案的样本时间相比,在延长照明条件(LL和DD)下维持的成年人的结果没有差异,后者存在明显的统计差异。胎儿视网膜的数据显示,在任何测量变量中都没有显著的可持续模式。这些定量研究结果得出结论,虽然周长测量的变化可以用囊泡循环假说来解释,但观察到的末端大小和形状的变化可能是由光激发或光增强机制控制的,并通过在这些光感受器突触末端发现的交叉条纹原纤维的环状结构来实现。
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引用次数: 4
In situ localization of lectin-binding glycoconjugates in the matrix of growth-plate cartilage. 生长板软骨基质中凝集素结合糖的原位定位。
Pub Date : 1986-05-01 DOI: 10.1002/aja.1001760106
C E Farnum, N J Wilsman

In the distal hypertrophic zone of growth-plate cartilage, the pericellular matrix surrounding individual chondrocytes and the territorial matrix uniting chondrocytes into columnar groups are invaded by metaphyseal endothelial cells prior to osteogenesis. In the present study, lectin-binding glycoconjugates were analyzed in these two matrix compartments of growth-plate cartilage from Yucatan swine. Nine lectin-fluorescein conjugates were tested by a postembedment method on 1-micron-thick, nondecalcified, Epon-embedded sections. Chondrocytes in all cellular zones were surrounded by a pericellular matrix which showed positive binding for peanut agglutinin (PNA), ricin agglutinin (RCA-I), and soybean agglutinin (SBA). Binding by these lectins was sensitive to digestion with hyaluronidase, chondroitinase, and trypsin. Pericellular glyconconjugtes that bind RCA-I and concanvalin A (CONA) after periodic acid oxidation, and which were sensitive to trypsin but not to chondroitinase or hyaluronidase, were present in the hypertrophic cell zone. Within the territorial matrix, binding of lectins specific for galactose, N-acetylgalactosamine, and fucose showed gradients of intensity which became maximal at the last transverse septum. Lectin-binding histochemistry more precisely differentiated the microheterogeneity of glycoconjugate distribution within these two matrix compartments than has been possible with other histochemical techniques. Lectin-binding affinity is a potentially useful technique by which to isolate cartilage matrix macromolecules unique to specific cellular zones of the growth plate.

在生长板软骨的远端肥大带,单个软骨细胞周围的细胞周基质和将软骨细胞联合成柱状群的区域基质在成骨之前被干骺端内皮细胞侵袭。本研究在尤卡坦猪生长板软骨的这两个基质室中分析了凝集素结合糖缀合物。9种凝集素-荧光素偶联物在1微米厚、非脱钙、epon包埋切片上采用包埋后方法进行检测。所有细胞带的软骨细胞都被细胞周基质包围,其与花生凝集素(PNA)、蓖麻毒素凝集素(rca - 1)和大豆凝集素(SBA)结合呈阳性。这些凝集素的结合对透明质酸酶、软骨素酶和胰蛋白酶的消化很敏感。细胞周围糖偶联物在周期性酸氧化后结合rca - 1和CONA (CONA),对胰蛋白酶敏感,但对软骨素酶或透明质酸酶不敏感,存在于肥大细胞区。在区域基质内,半乳糖特异性凝集素、n -乙酰半乳糖胺特异性凝集素与病灶的结合呈现强度梯度,并在最后横隔处达到最大。与其他组织化学技术相比,凝集素结合组织化学更精确地区分了这两个基质室中糖缀合物分布的微观异质性。凝集素结合亲和是一种潜在的有用的技术,通过它分离软骨基质大分子独特的特定细胞区域的生长板。
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引用次数: 19
The effects of orchidectomy and replacement therapy on the ultrastructure and gonadotropin-releasing hormone content of the median eminence of the rat. 兰花切除及替代疗法对大鼠正中隆起超微结构及促性腺激素释放激素含量的影响。
Pub Date : 1986-05-01 DOI: 10.1002/aja.1001760104
W C Beckman

The effects of 2 weeks of orchidectomy and replacement therapy with testosterone upon the content and distribution of gonadotropin-releasing hormone (GnRH) in the median eminence were determined by means of radioimmunoassay and electron microscopy. Photographic montages were prepared from electron micrographs of the lateral median eminence at the point of deepest invagination of the tuberoinfundibular sulcus. Morphometric analysis of photographs of tissues immunohistochemically stained for GnRH was performed to determine changes in the volume density of GnRH-containing axon profiles following the experimental treatments. A decrease in GnRH content after orchidectomy was observed both by morphometric analysis of axon volume density and radioimmunoassay of total GnRH content. Testosterone treatment of orchidectomized animals prevented the postorchidectomy loss of GnRH. Morphometric analysis of conventional electron micrographs revealed an increase in the number of axons containing no dense-core vesicles following orchidectomy, but no decrease in volume density of the neuropil. The results indicate that the change in volume density of immunostained axons was related to the loss of immunostainable dense-core vesicles and not to a change in the size or number of axons. The area corresponding to the location of the highest concentration of GnRH-containing axons was observed to be largely avascular and separated from the vessels of the tuberoinfundibular sulcus by a "border zone" composed of glial foot processes. The unique morphology of the GnRH area has suggested the name "compact zone" to distinguish it from the palisade zone with which it is continuous medially. GnRH axons in this region are probably part of a tract extending farther caudally rather than a terminal field.

采用放射免疫法和电镜观察睾丸激素替代治疗2周后对正中嵴促性腺激素释放激素(GnRH)含量和分布的影响。摄影蒙太奇是由电子显微照片的外侧正中隆起在最深的结节眼底沟内陷点。对免疫组织化学染色的组织照片进行形态计量学分析,以确定实验处理后含GnRH的轴突剖面体积密度的变化。通过轴突体积密度的形态计量学分析和总GnRH含量的放射免疫分析,观察到睾丸切除术后GnRH含量的下降。睾丸激素治疗可防止睾丸切除后GnRH的丢失。常规电镜形态计量学分析显示,睾丸切除术后不含致密核囊泡的轴突数量增加,但神经细胞的体积密度没有下降。结果表明,免疫染色的轴突体积密度的变化与免疫染色的致密核囊泡的丢失有关,而与轴突的大小和数量的变化无关。含有gnrh的轴突浓度最高的位置所对应的区域大部分是无血管的,由胶质足突组成的“边界区”与结节眼底沟的血管隔开。GnRH区域的独特形态建议将其命名为“紧凑区”,以区分它与连续的栅栏区。这个区域的GnRH轴突可能是更远的尾端延伸的束的一部分,而不是终端区。
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引用次数: 3
Factors that determine the form of neuromuscular junctions of intrafusal fibers in the cat. 决定猫肠内纤维神经肌肉连接形式的因素。
Pub Date : 1986-05-01 DOI: 10.1002/aja.1001760108
J Kucera, J M Walro

The form of terminations of fusimotor (gamma) and skeletofusimotor (beta) axons on intrafusal fibers was analyzed in serial sections of 20 spindles of the cat tenuissimus muscle. Seven synaptic features were assessed either qualitatively or quantitatively from electron micrographs of transverse sections of 184 intrafusal and 30 extrafusal endings. Features were compared among endings that were terminations of gamma or beta axons on different types of intrafusal fiber at different distances from the spindle equator. These comparisons indicated that interactions of several factors, and not the motor axon alone, determine the form of motor endings. Intrafusal muscle fiber type is dominant to the motor axon in regulation of the number and depth of postsynaptic folds. Separation of the influence of the motor axon from the muscle fiber was less clear with respect to the size of ending. Complete expression of the muscle fiber-motor axon interaction reflected by the form of motor endings is dependent upon location of the ending relative to the sensory region. Both depth of the primary synaptic cleft and size of the soleplate of motor endings increased with increasing distance of the ending from the spindle equator. A system of classification of cat intrafusal motor endings that reflects the multiplicity of factors that determine the form of endings, and one that simplifies the current terminology, is proposed.

在猫腱肌20个纺锤体的连续切片上分析了束内纤维上梭状运动(γ)和骨梭状运动(β)轴突的终止形式。通过对184个灌胃末端和30个灌胃末端横切面的电子显微图定性或定量地评估了7个突触特征。比较了不同类型的灌注纤维在离纺锤体赤道不同距离处的γ或β轴突末端的特征。这些比较表明,多种因素的相互作用,而不仅仅是运动轴突,决定了运动末梢的形式。在突触后褶皱的数量和深度的调节中,肌纤维类型对运动轴突起主导作用。运动轴突对肌纤维影响的分离在末端的大小方面不太清楚。肌肉纤维-运动轴突相互作用的完全表达是由运动末梢的形式所反映的,这取决于末梢相对于感觉区的位置。初级突触间隙的深度和运动末端底板的大小随运动末端离纺锤体赤道距离的增加而增加。本文提出了一种猫脑内运动末梢的分类系统,该系统反映了决定末梢形式的多种因素,并简化了当前的术语。
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引用次数: 16
Processing of concanavalin A-receptor complexes by rat osteoclasts in vitro. 大鼠破骨细胞体外加工刀豆蛋白a受体复合物的实验研究。
Pub Date : 1986-05-01 DOI: 10.1002/aja.1001760105
S N Popoff, G B Schneider

The osteoclast is a large multinucleate cell that is widely accepted as the primary effector cell responsible for normal bone resorption. In a previous study, we demonstrated that concanavalin A (con A) has a dose-dependent biphasic effect on the bone-resorbing capacity of osteoclasts, using a 45Ca bone-organ culture system; bone resorption was stimulated at low concentrations and inhibited at high concentrations. The mitogenic property of con A in lymphocyte cultures is well documented; therefore con A has been used extensively to study the manner in which lymphocytes and other mononuclear cells process the cell-bound lectin. In this study, we have investigated the processing of con A-receptor complexes by osteoclasts in culture, using con A-FITC to evaluate the redistribution of cell-bound con A via epifluorescence microscopy and using con A-ferritin to determine whether the lectin receptor complexes are internalized. The osteoclasts were obtained from the long bones of newborn rats and allowed to attach to glass coverslips at 37 degrees C. Following attachment, the nonadherent cells were removed by rinsing. The adherent osteoclasts were preincubated in 50 micrograms/ml con A-FITC or con A-ferritin at 4 degrees C for 10 min, washed to remove unbound con A, and incubated at 37 degrees C for 15 or 30 min in the absence of con A. Positive controls were fixed immediately after preincubation at 4 degrees C; negative controls were preincubated in con A-FITC and alpha-methyl mannoside, the haptenic inhibitor of con A binding. The results demonstrate that redistribution and endocytosis of con A-receptor complexes occurs within 30 min. These findings confirm the hypothesis that cell-bound con A can alter the structure and activity of osteoclast membrane components in a manner similar to that observed in mononuclear cell cultures. The internalization of con A may be important in altering osteoclastic activity by mediating intracellular mechanisms involved in the bone-resorbing process.

破骨细胞是一种大的多核细胞,被广泛认为是正常骨吸收的主要效应细胞。在之前的一项研究中,我们使用45Ca骨器官培养系统证明了刀豆蛋白a (con a)对破骨细胞的骨吸收能力具有剂量依赖性的双相效应;低浓度刺激骨吸收,高浓度抑制骨吸收。con A在淋巴细胞培养中的有丝分裂特性已得到充分证明;因此,con A已被广泛用于研究淋巴细胞和其他单核细胞处理细胞结合凝集素的方式。在本研究中,我们研究了破骨细胞在培养过程中对con A受体复合物的加工,使用con A- fitc通过荧光显微镜评估细胞结合的con A的重新分配,并使用con A-铁蛋白来确定凝集素受体复合物是否被内化。从新生大鼠的长骨中获得破骨细胞,并将其附着在37℃的玻璃罩上。附着后,通过冲洗去除非粘附细胞。将粘附的破骨细胞在50微克/毫升的con A- fitc或con A-铁蛋白中4℃预孵育10分钟,洗涤去除未结合的con A,在无con A的情况下37℃孵育15或30分钟,4℃预孵育后立即固定阳性对照;阴性对照在con A- fitc和α -甲基甘露糖苷(con A结合的半抗原抑制剂)中预孵育。结果表明,con - A受体复合物的重新分配和内吞作用在30分钟内发生。这些发现证实了细胞结合的con - A可以改变破骨细胞膜成分的结构和活性的假设,其方式类似于在单核细胞培养中观察到的。con A的内化可能通过介导参与骨吸收过程的细胞内机制,在改变破骨细胞活性方面发挥重要作用。
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引用次数: 5
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American Journal of Anatomy
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