人胚胎睾丸生殖细胞发育的组织学和超微结构研究

用光镜与电镜技术，对１５例７至２８周人胚胎睾丸生殖细胞的发育进行了研究。结果：（１）据时间，位置及形态结构特点，将生殖细胞的发育过程初步分为三个阶段，即原始生殖细胞阶段，生殖母细胞阶段和前的细胞阶段。其中前精原细胞阶段又经历了早，中，晚三上时期各期出现的时间有交错。（２）１４周是胚胎生殖细胞发育的一个重要时期。     

Prespermatogenesis and spermatogoniogenesis in the bovine testis

The bovine male germ cell population was studied over the entire period from testicular differentiation in the embryo through onset of spermatogenesis in the pubertal calf. Germ cells were identified by protein gene product 9.5 immunohistochemistry and characterized by their ultrastructure. The proliferation pattern of germ cells was studied with immunohistochemical anti-Ki 67 and anti-proliferating cell nuclear antigen reactions. Germ cells with a high proliferation rate are observed from day 50 p.c. to day 80 p.c. These cells are in transition from primordial germ cells to prespermatogonia. From day 80 p.c. until approximately the 15th postnatal week the germ cells present are identified as prespermatogonia. From day 80 p.c. to day 200 p.c. germ cell multiplication decreases continuously; then the prespermatogonia enter a phase of relative mitotical quiescence that lasts until the 4th postnatal week. Between the 4th and the 15th postnatal week, testicular tubular diameters grow from 40 to 80 μm and the prespermatogonia resume their proliferation. In seminiferous tubules with diameters between 80 and 120 μm, found in animals between 18 and 27 weeks of age, a central lumen is normally still absent. During this period germ cell proliferation reaches a second maximum. The cells involved represent the members of the spermatogonia stem and precursor cell line kinetically interpolated between the prespermatogonia and the first differentiating A-spermatogonia. This second phase of prepubertal germ cell multiplication coincides with the period when the pre-Sertoli cells transform into adult-type Sertoli cells and enter the G₀-phase for the rest of life. Accepted: 8 May 2000     

PROLONGED EXPRESSION OF THE c-kit RECEPTOR IN GERM CELLS OF INTERSEX FETAL TESTES

Stem cell factor (SCF) and its receptor Kit encoded by the c- kit proto-oncogene are crucial for the development and migration of primordial germ cells in rodents. The expression of Kit has been examined immunohistochemically in gonads obtained from five specimens of fetal tissues with intersex conditions which included 45,X/46,XY mosaicism; androgen insensitivity syndrome; and 46,XY/iso(p)Y mosaicism. Individuals with such disorders of sexual differentiation and Y-chromosome material carry a very high risk of developing testicular neoplasms. Fetal testicular germ cells of the intersex subjects expressed Kit at a later developmental age than controls, in which no Kit protein was detectable beyond the 15th week of gestation. This finding may indicate a disturbance of the chronology of germ cell development, or it may suggest a change of the regulation of c- kit expression in subjects with disorders of gonadal development.     

The liver hemopoietic environment: I. Developing hepatocytes and their role in fetal hemopoiesis

Mouse fetal liver was studied ultrastructurally to identify and characterize the developing hepatic parenchyma or prehepatocyte which may be responsible for producing the liver hemopoietic environment. It was observed that as the liver develops, there is close association of endodermal and mesenchymal cells in the region of the septum transversum. Numerous intercellular adhesions were observed between endodermal cells and mesenchymal cells. Twenty-four after endodermal and mesenchymal cells first intermingle, the liver extravascular space consisted of spherical hemopoietic cells dispersed among a heterogenous population of dark and light cells. The reticulum of prehepatocytes formed a three-dimensional cellular network which structurally supported the hemopoietic cells residing in the liver. By 12 days of gestation, prehepatocytes were a homogenous population of dark, stellate cells joined together by numerous intercellular adhesions. Broad areas of intercellular association were noted between processes and prehepatocytes and hemopoietic cells; however, no intercellular junctions between these two disparate cell populations were observed at this or any stage in development. Characteristics reflecting a cell population capable of synthesis and secretion of proteinaceous substances, namely, dilated Golgi apparati, increased numbers of polyribosomes and profiles of rough endoplasmic reticulum (RER), two types of vacuoles and/or vesicles, and intercellular microvillus-lined spaces, were observed in the prehepatocytes between 12 and 17 days gestation. By day 17 of gestation, glycogen accumulation, biliary channel development, appearance of a subendothelial microvillus surface, nuclear shape and chromatin pattern, and arrangement of cytoplasmic organelles reflected the maturation of prehepatocytes into hepatocytes, the adult liver parenchyma.     

Evolution of mesenchymal cells in fetal rat lung.

The evolution of connective tissue cells in the developing fetal rat lung is studied under the electron microscope from the 15th until the 21st day of gestation and is compared to the evolution of epithelial cells. Three successive types of stem cells ("mesocytoblasts") are present during the first stages of lung development studied (15 to 18 days of gestation). These stem cells appear to be able to differentiate into fibroblasts or into smooth muscle cells, according to their localization along the broncho-alveolar tubule. Myoblasts are situated near the bronchial epithelium, whereas fibroblasts occur under the alveolar epithelium. Epithelo-mesenchymal interactions are assumed to play a role in this differentiation process. Synthesis of both, collagen and elastic fibers and of cytoplasmic filaments by fibroblasts as well as by myoblasts reveal the multiple potentialities of the mesenchymal stem cell and suggest a common origin. The early fibroblast in characterized by long cytoplasmic processes which contain numerous cytofilaments, and by the presence of collagen fibers in the vicinity of the cell. Later on, (20 days of gestation) the mature fibroblast of the lung mesenchyme shows areas of RER, glycogen and lipidic vacuoles in its cytoplasm. Cytofilaments are numerous within very long cytoplasmic processes and elastic and collagen fibers are very frequent beside the cytoplasmic membrane. The earliest fibroblast differentiation occurs under the epithelium of primitive respiratory bronchioles, which indicate the limit between the bronchial and the alveolar territories. Later on, differentiating fibroblasts are found throughout the whole alveolar walls. Connective tissue cells other than mesenchymal stem cells, fibroblasts or myoblasts are observed during lung development. Vacuolar cells, similar to Hofbauer cells, transiently appear on the 16th day of gestation. On the 20th and the 21st day macrophage-like cells are present in the septal space of the alveolar wall. The absence of intermediate stages of differentiation and parallel evolution of blood cells suggest that those connective tissue cells are differentiated elsewhere and have then migrated from blood into lung mesenchyme. No cell death has been observed in the developing lung.     

An ultrastructural morphometric study of developing rat substantia gelatinosa

A morphometric analysis has been done on developing rat substantia gelatinosa of the lower cervical and upper thoracic levels of the spinal cord starting on the 15th day of gestation. The following parameters were measured: cell body diameter, cytoplasmic/nuclear areas, synaptic density, synaptic type and vesicle morphology of the presynaptic terminal in axodendritic synapses. Cell body size and cytoplasmic/nuclear areas of gelatinosal cells increase until the 15th day postnatally and then decrease somewhat to the adult values. The first synapses are seen on gestation day 17. Synaptic density increases linearly until the third day postnatally. Axodendritic synapses are most common throughout development and in the adult, while the proportion of axoaxonic synapses increases and axosomatic synapses decreases during development. Most of the terminals in axodendritic synapses contain clear-spherical vesicles but the occurrence of clear-flat vesicles and dense-cored vesicles in the terminals increases during development. It appears that these morphological parameters provide a stable index of development in the substantia gelatinosa which can be correlated with functional development of the area. Hopefully, they will provide a means to assess subtle anomalies induced by nonteratogenic drugs or other environmental changes.     

人胎睾丸的组织发生

用7～38周人胎35例,取睾丸固定于 Carnoy 液,石蜡包埋,以 HE、PAS 反应及甲绿哌喏宁法染色。7周初的性腺尚未分化,可见散在的原始生殖细胞。13周睾丸特征已明显,白膜厚,睾丸素清楚,索间有密集的嗜酸性间质细胞。14.5周部分睾丸索出现小腔,而成管状。睾丸索包含大而着色淡的原始生殖细胞及色深的原始支持细胞。胎早期原始生殖细胞含有丰富的糖原颗粒。睾丸门、白膜及表面上皮内均见散在或成群的生殖细胞。睾丸间质细胞分幼稚型、成熟型及退化型。13～15周以幼稚型为主;16～18周以成熟型为主,成熟型的胞质内含有 RNA 颗粒;20周后退化型增多,成熟型减少。38周时,睾丸间质细胞单个或成行存在,数量大为减少。     

Oct-4和人端粒酶逆转录酶在不同胎龄人胚原始生殖细胞的表达变化

目的 检测不同胎龄人胚原始生殖细胞中Oct-4、人端粒酶逆转录酶(hTERT)的表达变化.方法 取人胚胎生殖嵴,通过RT-PCR检测5～13周龄人胚生殖嵴中Oct-4、hTERT的表达变化;同时,取人胚胎生殖嵴,经石蜡切片,HE染色、免疫组织化学染色等技术,观察不同胎龄人胚原始生殖细胞的形态,检测Oct-4、hTERT的表达情况.结果 胎龄6～7周时,生殖嵴中可见少量原始生殖细胞,且表达Oct-4及hTERT;胎龄8～12周时,生殖嵴中原始生殖细胞数量增多,Oct-4及hTERT表达增强(P<0.05);胎龄13周以后,生殖嵴中Oct-4阳性的原始生殖细胞数量逐渐减少,Oct-4表达减弱,但hTERT仍然高表达.结论 不同胎龄人胚原始生殖细胞中Oct-4的表达呈动态变化,胎龄8～12周时表达较强,但hTERT的表达量始终维持在较高水平,没有明显变化.     

Development of type I cells of the rabbit subclavian glomera (aortic bodies): A light,fluorescence and electron microscopic study

The embryogenesis of the subclavian glomera (aortic bodies) is controversial. Past investigators have attributed the development of the Type I cells to mesodermal and/or neural elements. Based on the results of the present light microscopic, fluorescence histochemical and electron microscopic study of rabbit aortic bodies from 16 days of gestation (term: 31 days) to four days postpartum, it appears that the Type I glomus cells are derived from cells of neural crest origin. The subclavian glomus anlage is associated with cells of vagal origin throughout its development. Evidence of Type I cell development from pre-existing mesodermal condensations is not observed. Type I cells exhibit formaldehyde-induced-fluorescence by the twentieth day of gestation. Dense-cored cytoplasmic vesicles are apparent by the sixteenth day of gestation. The number of cytoplasmic vesicles increases steadily, but the greatest increase of vesicles is observed between the twenty-eighth day of gestation and birth. Primitive Type I glomus cells exhibit abundant polysomes and rough endoplasmic reticulum indicative of synthetic activity. Nerve terminals are apparent adjacent to Type I cells by the twentieth day of gestation, but synaptogenesis does not occur until sometime between the twenty-fourth and twentyeighth days of gestation. Abundant vascularity, characteristic of chemosensory glomera, is not achieved until the twenty-eighth day of gestation.