应用多重PCR技术检测性器官异常患者的性别决定基因

采用针对人SRY基因及X与Y染色体同源序列的两对引物进行多重聚合酶链反应技术检测204例新生儿脐血DNA,男性均显示590bp、 355bp及280bp 3条扩增带,女性只有590bp1条扩增带,性别鉴定准确率为100%。又检测47例性器官异常患,13例社会性别为女性者只出现590bp扩增带,与核型性别结果一致。 34例社会性别为男性者中,有2例SRY基因检测结果阴性,只出现590bp带,核型为46,XX,证明这两例患者社会性别不相符,经病理证实后应诊断为女性假两性畸形。 Abstract:This investigation adapted multiplex PCR technique with two pairs of primer to determine sex according to the human SRY gene and X,Y chromosome analogical sequence.We detected bellybutton blood DNA from 204 newborns.Male revealed three bands:590bp,355bp and 280bp,and females only had one band which was 590bp,the accuracy of sex determination was 100%.47 sexual abnormal patients were tested.The results showed that 13 cases,whose social sex were female,had one 590bp band which consistent with their chromosome type sex.In the rest 34 cases,whose social sex were male,2cases showed,on the contrary,only one 590bp band and their chromosome type were 46,XX.This proved that the two patients did not consistent with their social sex,and pathological analysis showed that they were female pseud-hermaphroditism.     

利用差减杂交分离具有进化保守性的睾丸表达的基因

The mechanisms of sexual development and regulation display extraordinary diversity between phyla. However, recent studies show that there must be underlying evolutionary conservatism between different mechanisms. In order to examine evolutionary conservatism in molecular mechanisms of sex determination, the natural sex reversal process of the rice field eel (Monopterus albus) was used to identify conserved functional genes involved in sex determination and differentiation. Firstly, a mini cDNA library was constructed from the cDNAs expressed specifically in the human testis. The cDNA library was screened with first strand cDNA probes from the intersex gonads of rice field eels. Secondly, likely positive clones were arrayed onto two nylon membranes, which were then hybridized with the first strand cDNAs probes from rice field eel intersex and female gonads respectively. Two positive clones were obtained, which showed obvious differential expression between intersex and female gonads. DNA sequencing analysis reveal that the insert fragments of these two clones were 895 bp and 596 bp long respectively. BLAST results indicate that the 895 bp fragment is the 3′ part of the No.AJ011779 gene, located on 6q21, encoding a homologue of yeast Sec63p protein. An interesting finding is that one member of the human Sec gene family, Sec61γ , is expressed in the testis and involved in the process of testis development. The 596 bp fragment was 97% identical with the No.BG723880 EST, located on clone RP11 12M5 of chromosome 1 This needs further study. We infer that these two genes should also play a role in sexual development.     

黑麂Y染色体的鉴别和Sry基因的克隆及定位

以流式细胞仪分离小麂(Muntiacus reevesi)Y染色体和黑麂(Muntiacus crinifrons)Y1,Y2,X+4和1号染色体,利用DOP-PCR技术富集了分离的各单条染色体。然后,将小麂的Y染色体的DOP-PCR产物经Cy3标记后直接作为涂染探针,应用染色体涂染技术与雌雄黑麂的核型标本进行杂交,确认了黑麂真正的Y染色体为Y2染色体。再以黑麂的Y1,Y2,X+4和1号染色体的DOP-PCR产物为模板,用人的特异性的SRY（sex determining region of the Y chromosome）基因引物对其进行扩增,结果表明黑麂只有Y2染色体出现了SRY扩增片段。然后扩增产物克隆和测序,比较它与人的同源性,初步把黑麂的Sry基因定位在Y2染色体上。最后提取雄性黑麂的基因组DNA,并用同一对引物对其进行扩增,亦得到Sry基因的片段,对此扩增片段进行克隆,测序,结果表明其与Y2染色体得到的Sry基因片段完全一样,与人SRY基因的同源性均为83%。 Abstract：The single Y chromosome of Muntiacus reevesi and Y1,Y2 ,X+4,1 chromosome of Muntiacus crinifrons were obtained by flow-sorting ,then they were amplified through DOP-PCR . After that, the metaphase karyotype of Muntiacus crinifrons were painted by using the product of the DOP-PCR of the Y chromosome of Muntiacus reevesi as a special probe and the result showed that Y2 chromosome was the real Y chromosome of Muntiacus crinifrons. Secondly the product of the DOP-PCR of Y1,Y2,X+4,1 chromosome of Muntiacus crinifrons were used as the templates of the next amplification using the special primer devised according to the human SRY gene .One band was obtained only from Y2 chromosome, then it was cloned to the T-vector and sequenced. The Sry gene sequence of Muntiacus crinifrons was acquired and the conclution was that there are 83% homology between the human and Muntiacus crinifrons. It was testified that in all mammal Sry gene is consertive. On the other side the Sry gene was located to the Y2 chromosome of the Muntiacus crinifrons.     

羊驼染色体核型及其G分带的研究

为了从选种、杂交改良、疾病诊断以及性别决定的遗传机制等方面为羊驼的繁育与推广提供更为有效的细胞遗传学资料,本试验采用外周血淋巴细胞培养法及胰酶-EDTA法分析了23只胡阿基亚型羊驼（Huacaya alpaca,雌20只,雄3只）的染色体核型及其G-分带,结果表明：羊驼二倍体染色体数目为2n=74,雄性羊驼核型为74,XY;雌性羊驼核型为74,XX。其中,1～20对常染色体为亚端着丝粒染色体,21～36对常染色体为亚中着丝粒染色体和中着丝粒染色体,X为中着丝粒染色体,Y为端着丝粒染色体。G-带分析表明,羊驼G带明暗相间,显现出不同的带纹,且羊驼每对染色体都有其独特的带纹特征,其带纹数目和精细程度随着染色体长度的增加而增加。Abstract: Blood samples from 23 Huacaya alpacas, 3 males and 20 females, were used to study chromosomes and karyotypes, so as to provide some effective cytogenetic bases for the selection, improvement by crossing, disease diagnosis of alpacas, and genetic mechanisms of sex determination. Peripheral blood lymphocyte culture was used to prepare chromosome. A method of trypase-EDTA was used for G-banding. The results showed as follows: The number of diploid chromosomes was 2n=74, with the karyotype 74, XY and 74, XX for males and females respectively. Thirty-six homologous pairs of chromosomes were autosomes, in which chromosomes pairs No.1 to No.20 were acrocentric-subterminal and No.21 to No.36 metacentric-submetacentric. And X chromosome was metacentric, Y chromosome telocentric. The analysis of G-bands showed that bright and dark bands appeared by turn. It showed different bands. And every pair of chromosomes had its distinct band, and the longer the chromosomes, the more the number of bands, and the more clear the bands.     

Variation in chromosome number in the seedling progeny of a somaclone of Paspalum dilatatum

The somaclone,C39,derived by tissue culture from the obligate apomict Paspalum dilatatum cv Raki(2n=50),had 50 chromosomes and a karyotype apparently identical to Raki.SC2 seedlings of C39 showed a high degree of phenotypic variation which was often associated with increased chromosome numbers,but some of the variant seedlings were karyotypically indistinguishable from Raki or C39.Plants with increased chromosome numbers exhibited a high degree of intraplant chromosome variation(aneusomaty).In one of the SC2 seedlings,the chromosome number of root tip cells varied from 58 to 82 and in several other seedlings the range was more than 10.The results suggested that the ability to form seed apomictically was much reduced in C39 and that this plant showed some capacity for sexual reproduction and the resulting seedlings,with a chromosome number of about 70,were genetically unstable.Of 11 SC2 seedlings examined cytologically,6 did not produce any viable seed.Seedlings grown from seed of the remaining 5 plants showed that aneusomaty persisted in the SC3 generation.SC3 seedlings which were phenotypically similar to their maternal parent showed a similar range of chromosome numbers to that parent.Some of the SC3 seedlings exhibited an even wider range of chromosome numbers(e.g.56-136),and these plants were all dwarfs.     

Sex differences in morphine-induced behavioral sensitization and social behaviors in ICR mice

Gender and genetic strain are two prominent variants that influence drug abuse. Although certain sexrelated behavioral responses have been previously characterized in ICR mice, little is known about the effects of sex on morphine-induced behavioral responses in this outbred strain. Therefore, in this study, we investigated the sex differences of morphine-induced locomotion, anxiety-like and social behaviors in ICR mice. After morphine or saline exposure for four consecutive days(twice daily), increased locomotion, more time spent in the central area, as well as attenuated rearing and self-grooming behaviors were found in morphine-treated females in an open field; no differences were found in locomotion and the time spent in the central area between male and female controls. When interacting with the samesex individuals, female controls were engaged in more social investigation, following, body contacting and self-grooming behaviors than controls; morphine exposure reduced contacting and self-grooming behaviors in females; in contrast, these effects were not found in males. These results indicate that female ICR mice are more prosocial and are more susceptible to morphine exposure than males.     

大鳞副泥鳅ZZ/ZW型性别决定的细胞遗传学证据

大鳞副泥鳅是鲤形目、鳅科的鱼类。其2n数为48,核型组成为12m+4sm+32 t(雄性),11m+5sm+32t(雌性)。根据银染带和C带特征分析,证实大鳞副泥鳅为ZZ/ZW型性别决定。Z染色体为中部着丝粒染色体,在其长臂端部有Ag -NOR存在。 W染色体为亚中部着丝粒染色体,在其长臂末端也有Ag-NOR存在,同时还有一深染的居间C带,这是W染色体独有的带纹特征。 Abstract:Paramisgurnus dabryanus belongs to Cypriniformes,Cobitidae.Its 2n is 48.The karyotype formula is 12m+4sm+32t(in male),11m+5sm+32t(in female).According to the Ag-NORs band and C-band patterns,we consider that its sex determination is of ZZ/ZW type.The Z chromosome is a metacentric one with Ag-NORs located on its arm end.The W chromosome is a submetacentric with Ag-NORs located on the terminal of its long arm.There is a darkly stained C-band on the long arm of W chromosome.This band is a characteric of the W chromosome.     

Reconsidering sex differences during place learning in tungara frogs

In tungara frogs, female mate choice requires remembering the location and/or calls of preferred males who advertise from fixed positions within a breeding pond. A previous study found that, when solving a place discrimination task in the laboratory, female tCingara frogs were able to learn a visual cue to solve the task, whereas males were not. In that task, male performance appeared to be inhibited, in part, by their attempt to use egocentric cues. We tested whether the sex difference in place learning previously reported would generalize to other training parameters with different cues available by eliminating the potential to use egocentric cues and increasing the number of trials per day. As before, frogs were given a choice between a red or yellow door, one of which led to shelters and return to their home cage. In the current testing conditions, we detected a preference for the red door;thus, we only considered frogs rewarded to the yellow door. Training was associated with an in crease in correct choices and an in creased preference for the yellow door. However, there was no evide nee for a sex d iff ere nee in learni ng. In summary, un der the curre nt training conditions, we fou nd that the appare nt female advantage in place learning was no Ion ger evide nt. Future studies that in vestigate sex d iff ere nces in cue pref ere nee and/or ability to switch among cues will further illuminate the conditions under which sex differences in learning are manifest in tungara frogs.     

Tumor suppressor gene PTEN affects anoikis via both PKB and FAK in human lung carcinoma cell lines

PTEN was identified in 1997 as a new tumor suppressor gene on human chromosome 10q23, deletions and mutations of this gene were associated with a variety of human cancers such as glioblastoma and prostate cancer. In this study Northern blot analysis was carried out and one major band at 2 kb region was observed in all 8 HLCC samples, consistent with previous reports. The result showed that the PTEN gene were expressed and its mRNA level similar in all cell lines tested. To determine whether the PTEN mRNA level reflects the parallel level of protein, the level of PTEN protein was examined by Western blot.PTEN protein level was high in H460 and detectable in A549, A4, A7, L1 cells, not detectable in 95C, 95D, A1     

泰和乌骨鸡的核型与带型研究

采用外周血淋巴细胞培养--空气干燥法,对泰和乌骨鸡染色体核型和带型进行了研究。结果表明：泰和乌骨鸡体细胞染色体数目2n=78,染色体基本臂数AF=90,1、9号染色体及Z、W性染色体为中央着丝粒（m）染色体,2、4、7号染色体为亚中央着丝粒（sm）染色体,3、6、8、10号染色体为端着丝粒（t）染色体。G带研究表明：前10对大型染色体可分为29个区,190条带。C带处理发现,所有母鸡分裂相中W性染色体都出现C带并整条深染。Ag-NORs研究发现：Ag-NORs常定位于1、2号常染色体短臂和Z性染色体短臂端部;Ag-NORs数目分布范围为1～6;平均每个细胞的Ag-NORs数在雌、雄鸡中分别为2.94和2.96。 Abstract:This study made the chromosome slides of Taihe Silkies by the peripheral blood lymphocyte culture-drying method,and analyzed Taihe Silkies karyotype and band pattern.The results are as follow:The diploid chromosome number of Taihe Silkies was 2n=78,the basic number of chromosome arms was AF=90 and the sex chromosome type was ZZ(♂)/ZW(♀).According to the measured relative length,arm ratio and centromeric index,the first 10 pairs of macro-chromosomes are described as follows:No.1 ,9 and Z,W chromosomes were metacentrics,No.2,4,7 were submetacentrics,and No.3,6,8,10 were telocentrics.Studies on Taihe Silkies′ G-band showed that the first 10 pairs of macro-chromosomes can be divided into 29 zones and 190 bands.Being treated by C-banding technique,a totally dark-stained and easily indentified W-chromosome always showed up in the female metaphase configurations.Ag-NORs were located in the short arms′ telomere of No.1,2 euchromosomes and Z sexchromosome,the Ag-NORs number varied from 1-6. -NORs     

Investigation of the mechanism of temperature sensitivity of the electroreceptors of ampullae of lorenzini

In experiments on Black Sea skates (Raja clavata), the potential of the receptor epithelium of the ampullae of Lorenzini and spike activity of single nerve fibers connected to them were investigated during electrical and temperature stimulation. Usually the potential within the canal was between 0 and –2 mV, and the input resistance of the ampulla 250–400 k. Heating of the region of the receptor epithelium was accompanied by a negative wave of potential, an increase in input resistance, and inhibition of spike activity. With worsening of the animal's condition the transepithelial potential became positive (up to +10 mV) but the input resistance of the ampulla during stimulation with a positive current was nonlinear in some cases: a regenerative spike of positive polarity appeared in the channel. During heating, the spike response was sometimes reversed in sign. It is suggested that fluctuations of the transepithelial potential and spike responses to temperature stimulation reflect changes in the potential difference on the basal membrane of the receptor cells, which is described by a relationship of the Nernst's or Goldman's equation type.I. P. Pavlov Institute of Physiology, Academy of Sciences of the USSR, Leningrad. I. M. Sechenov, Institute of Evolutionary Physiology and Biochemistry, Academy of Sciences of the USSR, Leningrad. Pacific Institute of Oceanology, Far Eastern Scientific Center, Academy of Sciences of the USSR, Vladivostok. Translated from Neirofiziologiya, Vol. 12, No. 1, pp. 67–74, January–February, 1980.     

Principles of organization and evolution of systems of regulation of functions

Evolution of living organisms is closely connected with evolution of structure of the system of regulations and its mechanisms. The functional ground of regulations is chemical signalization. As early as in unicellular organisms there is a set of signal mechanisms providing their life activity and orientation in space and time. Subsequent evolution of ways of chemical signalization followed the way of development of delivery pathways of chemical signal and development of mechanisms of its regulation. The mechanism of chemical regulation of the signal interaction is discussed by the example of the specialized system of transduction of signal from neuron to neuron, of effect of hormone on the epithelial cell and modulation of this effect. These mechanisms are considered as the most important ways of the fine and precise adaptation of chemical signalization underlying functioning of physiological systems and organs of the living organism