Morphological,histochemical and biochemical studies on germ cell mitochondria of normal rats

Summary Morphological changes in rat germ cell mitochondria are described. In diplotene and secondary spermatocytes and in the spermatids of the Golgi, cap and acrosomal phases, the mitochondria take on a rounded appearance with the inner space containing the matrix flattened against the outer membrane and the intracristal spaces considerably swollen (condensed mitochondria).Functional studies on condensed mitochondria isolated from the germ cells of normal rats have been performed. The following parameters have been evaluated: ADP/O ratio, respiratory control ratio (RCR) and ADP affinity. The ADP/O values found in the presence of various substrates are in agreement with the theoretical figures. The RCR is remarkably high. Moreover, the ADP affinity of these mitochondria is very high, as demonstrated by the low values of the apparent Km. These biochemical findings, which demonstrate a high oxidative capacity coupled with a marked phosphorylation, suggest that the condensed appearance of germ cell mitochondria is the expression of an active functional state.The work was partially supported by a grant from The Consiglio Nazionale delie Ricerche (C.N.R.), Rome, Italy     

原癌基因c-myc、c-H-ras、c-sis在小鼠卵巢和睾丸中的转录表达研究

以地高辛标记c-myc、c-H-ras、c-sis基因作探针,利用组织切片原位杂交的方法,检测了昆明种小鼠5周龄、10周龄和孕期卵巢,以及1天龄、5周龄和10周龄睾丸,初步研究了这3种原癌基因在生殖腺中的转录表达及其可能的功能作用。结果显示:(1)c-myc基因在孕鼠卵巢的个别卵泡的卵泡细胞中有强转录信号,但在3组卵巢的卵母细胞中及3组睾丸切片中均未见杂交信号。(2)c-H-ras基因在孕鼠卵巢黄体中有较强转录表达,但在3组卵巢的卵母细胞中及3组睾丸切片中均无杂交信号,(3)c-sis基因在10周龄睾丸曲细精管的精原细胞层和精母细胞有强转录表达,而在1天龄、5周龄睾丸及3组卵巢切片中均无杂交信号。上述结果表明:c-myc基因可能与精子发生和卵子的发育关系不大,但可能与卵泡的退化有关;c-H-ras基因可能与精子发生和卵子的发育关系也不大,但可能与妊娠黄体的形成有关;c-sis基因可能与卵子的发育无关,但可能与精子的发生密切相关。 Abstract:Using in Site Hybridization on tissue section of ovary from 5 weeks old,10 weeks old and pregnant Kunming mice,and test is from one-day old,5 weeks old,and 10 weeks old Kunming mice,we prelimmaily studied the expression and function of the proto-oncogenes c-myc,c-H-ras and c-sis.The results are as follows:(1)The strong expression of c-myc gene was found in follicular epithelium of some follicles of ovary from pregnant mice,and was not found in the oocytes of three groups of female mice and testes of three groups of male mice.(2)The strong expression of c-H-ras was observed in corpus luteum of ovary of pregnant mice,but it was not observed in any germ cells of all three groups of female mice and male mice.(3)The strong expression of c-sis gene was found in spermatogomia and spermatocytes of mature male mice,but it was not found in any germ cells of one day old and immature male mice and three groups of femala mice.These results showed that there might be no relationships between the expression of c-myc gene and spermatogenesis or oocyteatogenesis,but the expression of c-myc gene probably related to the cells degeneration of follicle;the expression of c-H-ras gene hadn’t a bearning on oogenesis and spermatogenesis,but probably related to the genesis of corpus luteum of corpus luteum of pregnant mice;the expression of c-sis gene there might be no related to the oogenesis and sperm maturation,but probably promoted the division of spermatogonim.     

Cathodoluminescence-emitting lipid droplets in rat testis: a study by analytical color fluorescence electron microscopy

Cathodoluminescence (CL) from lipid droplets (LDs) in the rat testis was examined by analytical color fluorescence electron microscopy. The results show that (1) the Cl at wavelengths of 320 nm (CL320) and 450 nm (CL450) is derived from cholesterol esters and a mixture of lipids including vitamin A esters, respectively; (2) CL320 in the LDs of Leydig cells sharply decreases on postnatal day 21, while CL320 and CL450 in the LDs of Sertoli cells begin to be detectable; (3) the CL450-emitting LDs in seminiferous tubules, whose distributional patterns display cyclic changes during the spermatogenic cycle, are involved in spermatogenesis; and (4) the intensity of CL as well as the distributional patterns of CL-emitting LDs in testicular cells change after hypophysectomy, vitamin-A deficiency, and treatment with ethylene dimethane sulfonate and testosterone propionate. This study demonstrates that analytical color fluorescence electron microscopy is a useful tool for in-vivo observation of some specific compounds which cannot be visualized by other methods.     

The shape and distribution of lysosomes and endocytosis in the ciliary epithelial cells of rats

Stages of the spermatogenic cycle in human seminiferous tubules were evaluated in men with varied efficiencies of spermatogenesis to determine if the architectural arrangement of stages or the atypical cell types contributed to variation in sperm production rates. Testes were selected from men with low, intermediate, and high daily sperm production per g parenchyma (DSP/g). Round tubular cross sections were photographed by bright-field microscopy. Stages were identified for each cross section by two observers and the number of stages represented in each cross section was recorded. Number of stages per cross section in men with low efficiency of spermatogenesis were significantly (P<0.05) fewer than men with intermediate and high efficiency of spermatogenesis. Further, the percentage of stages with atypical cell types in men with high DSP/g was significantly (P<0.05) higher than men with low DSP/g. There was a significant relationship (P<0.01) between the percentages of stages with atypical cell types per stage and number of stages per cross section. The atypical cell types appear to result from high density of stages per cross section in men with high DSP/g. There was no significant difference observed between groups for tubular volume, diameter, length, volume density, and volume density of seminiferous epithelium. However, a significant (P<0.05) positive correlation between percent seminiferous epithelium per testis with DSP/g or with the number of stages per cross section was found. These findings reveal that the architectural makeup of stages within seminiferous tubules and atypical cell types within stages varies with the level of efficiency of spermatogenesis, and this variation may reflect differences in yield of early spermatogonial divisions that are responsible for generating the different stages.     

国人睾丸、附睾和输精管的氧化还原酶的组织化学观察

本文收集了１９—３８岁国人正常男性新鲜睾丸、附睾和输精管１３例,进行了氧化还原酶组织化学染色、光镜定位及定性观察。结果表明：睾丸曲细精管和输出小管上皮的ＧＤＨ,ＮＡＤＨＤ,ＮＡＤＰＨＤ,ＳＤＨ,ＧＰＤＨ,ＩＣＤＨ,ＭＤＨ,ＬＤＨ和Ｇ－６－ＰＤＨ９种酶;睾丸间质细胞和附睾管上皮的ＮＡＤＨＤ,ＮＡＤＰＨＤ,ＳＤＨ,ＩＣＤＨ,ＭＤＨ,ＧＤＨ,ＬＤＨ和Ｇ－６－ＰＤＨ８种酶;输精管的ＮＡＤＨＤ,ＮＡＤＰＨＤ,ＩＣＤＨ和ＧＤＨ４种酶的酶活性呈强阳性或极强阳性。提示输出小管和头部附睾管含有的多种氧化还原酶对精子功能成熟有极重要作用。     

Expression of scatter factor/hepatocyte growth factor is regionally correlated with the initiation of sperm motility in murine male genital tract: Is scatter factor/hepatocyte growth factor involved in initiation of sperm motility?

Based upon findings that the scatter factor/hepatocyte growth factor (SF/HGF) has strong mitogenic and motogenic properties, and that the sperm cell acquires its fertilizing capacity and motility in the distal parts of mammalian epididymis, the present study was conducted to investigate the role of SF/HGF in initiation of sperm cell motility. This was investigated by determining the expression of SF/HGF in various regions of the murine male genital tract by scatter and cell tracking assays using MDCK epithelial cells, Western blot procedure, and the immunohistochemical procedure using paraffin sections of various regions of the male genital tract. The findings from all these assays indicate that SF/HGF is differentially expressed in various parts of the male genital tract with slight or no expression in the testes, caput epididymis, and vas deferens, and with the highest expression in cauda and corpus (distal) epididymis followed by expression in the corpus (proximal) epididymis. This region-specific SF/HGF expression pattern coincides with the pattern of acquiring the fertilizing capacity and motility by the sperm cell during its transit through the male genital tract. However, wherever SF/HGF was expressed in the male genital tract, its molecular weight was slightly higher (Mr, 82 kD), compared to the SF/HGF expressed in various other somatic tissues (Mr, 78 kD), indicating that the genital tract SF/HGF may be a different molecular species that shares some immunoreactive epitopes with the somatic cell SF/HGF. Incubation of immotile sperm from caput epididymis with the purified human placental SF/HGF of 78 kD initiated motility in 5–15% of sperm population. These results strongly suggest that the SF/HGF-like activity is expressed in the male genital tract in a region-specific manner, and this activity may have a role in initiation of sperm motility acquired during its transit through the epididymis in mammals. © 1994 Wiley-Liss, Inc.     

Temperature regulation of the testes of the bottlenose dolphin (Tursiops truncatus): evidence from colonic temperatures

Dolphins possess a countercurrent heat exchanger that functions to cool their intra-abdominal testes. spermatic arteries in the posterior abdomen are juxtaposed to veins returning cooled blood from the surfaces of the dorsal fin and flukes. A rectal probe housing a linear array of five copper-constantan thermocouples was designed to measure colonic temperatures simultaneously at positions anterior to, within, and posterior to the region of the colon flanked by the countercurrent heat exchanger. Colonic temperatures adjacent to the countercurrent heat exchanger were maximally 1.3°C cooler than temperatures measured outside this region. Temporary heating and cooling of the dorsal fin and flukes affected temperatures at the countercurrent heat exchanger, but had little or no effect on temperatures posterior to its position. These measurements support the hypothesis that cooled blood is introduced into the deep abdominal cavity and functions specifically to regulate the temperature of arterial blood flow to the dolphin testes.     

树鼩(TUPAIA BELANGERI CHINENSIS)精子发生的季节性变化

对野外成体树鼩进行了不同季节精小管发育的测定和生精上皮细胞学特征的观察。精小管平均直径的高峰值出现在4月,为205.7μm:最低值出现在10月,为88.4μm。1、4月份的生精上皮,表现了高度活跃的精子发生,具有明确周期的12个连续阶段和典型的细胞组合。生精上皮退行性变化最早出现于7月,8月进一步发展,10月最明显。附睾精子数量亦表现了类似的季节性波动。表明树鼩精子发生的活性期始于仲冬,最盛在春季,退行期始于仲夏,秋季最显著,有一个明显的精子发生的季节性周期。季节性生殖受局部环境因子的调节,光照可能是重要因素。     

Expression of fetal-type intermediate filaments by 17-day-old rat Sertoli cells cultured on reconstituted basement membrane

Summary The expression of cytokeratin- and vimentin-type intermediate filaments was studied by means of immunohistochemistry in Sertoli cells cultured on two types of reconstituted basement membrane in two-compartment culture chambers. In situ, the Sertoli cells of 17-day-old rats contained only vimentin intermediate filaments. During culture, a gradual reorganization of intermediate filaments accompanied by an increased cytokeratin immunoreactivity was observed. After 6 days, Sertoli cells contained both cytokeratin and vimentin, and the same cytokeratin type as in fetal and newborn testis was revealed by electrophoresis and immunoblotting. The present study shows that the isolation and culture of Sertoli cells causes, even in an improved culture system qualitative changes in the expression of intermediate filament proteins.