AIM To find out the relationship between the gene transcription of different types ofprocollagen and the deposition of the relevant collagens in the liver tissue and to confirm the types of collagen producing cells in...AIM To find out the relationship between the gene transcription of different types ofprocollagen and the deposition of the relevant collagens in the liver tissue and to confirm the types of collagen producing cells in liverfibrogenesis.METHODS Dynamic changes of the expression of α1(Ⅰ ), α1 (Ⅲ ) and α1 (Ⅳ) procollagen mRNAand relevant collagens and the distribution ofcollagen producing cells during liver fibrogenesis of rat induced by CCl4 (20 weeks)were investigated with Northern blot analysis,in situ hybridization and immunohistochemicaltechniques.RESULTS The increased expression of α1 (Ⅲ)procollagen mRNA by Northern blot analysis was the most predominant one among the threemRNAs during fibrogenesis. However, theenhanced expression of al (Ⅳ) procollagenmRNA occurred very early while the expressionof α1 (Ⅰ) mRNA was not enhanced much until themiddle stage of the exPeriment. D6smin (Dm)positive hepatic stellate cells (HSCs) and fewmyofibroblasts (MFs) in and around the necrotic areas expressed α1 (Ⅰ), α1 (Ⅲ) and α1 (Ⅳ)procollagen mRNA signals detected by in situhybridization at the early stage of theexperiment. All the three procollagen mRNAsignals thereafter mainly localized in fibroblasts (Fbs) and MFs in fibrotic septa during the middle and late stages of fibrosis, which distributedparallel to the corresponding collagens detected by immunohistochemical study. ln addition, the endothelial cells of sinusoids and the small blood vessels within the septa also showed α1(Ⅳ) procollagen mRNA and type Ⅳ collagen expressionCONCLUSION It is considered that "HSC-MF-Fb" effect cell system is the major cellularsource of collagen production in liver fibrosis, in which HSCs are collagen producing precursor cells in the early liver fibrogenesis, thereafter the synthesis of type Ⅰ, Ⅲ and Ⅳ collagens (Col Ⅰ, Col Ⅲ and Col Ⅳ) mainly derives fromMFs and Fbs, which play a very important role in the progress of liver fibrosis. The endothelialcells along sinusoids, as another source of Col 展开更多
Kainic acid (KA) is a structural analogue of glutamate with potent excitotoxic andconvulsant properties. Intraperitoneal injection of KA in rats produces recurrentspontaneous limbic seizures accompanied by brain damag...Kainic acid (KA) is a structural analogue of glutamate with potent excitotoxic andconvulsant properties. Intraperitoneal injection of KA in rats produces recurrentspontaneous limbic seizures accompanied by brain damage reminiscent of Ammon’s hornsclerosis, which has been considered as a useful model of human temporal lobe epilep-展开更多
Objective To explore specific gene expression for regulating meiosis of germ cells during spermatogenesis of rat testis Materials & Methods Male SD rats, aged 1, 3 and 8 weeks, were observed in this study. The ...Objective To explore specific gene expression for regulating meiosis of germ cells during spermatogenesis of rat testis Materials & Methods Male SD rats, aged 1, 3 and 8 weeks, were observed in this study. The methods of morphological observation on testicular tissues embedded by resin and mRNA differential display (DDRT PCR) were combined to obtain specific mRNA expression gene fragments during the testicular development. Reverse dot blot hybridization was operated to further screen the positive differential DNA fragments. The positive DNA segments were sub cloned in pGEM T Easy vector and transformed into the competent E coli 109 straint. Northern blot analysis and in situ hybridization were also carried out for identifying tissue specific expression as well as cell specific expression DNA fragments. To screen λ ZAP II rat testicular gene library was searched for the original gene. Results Eighty two differential cDNA fragments were obtained through primary DDRT PCR, among which 40 differential cDNA fragments were selected for further screening with reverse dot blot hybridization. After the reverse dot blot hybridization, 12 primary differential DNA fragments were obtained. The size of DNA fragments ranged from 250 to 500 bp. The in situ hybridization of the testicular tissue showed that a specific DNA fragment derived from 8 week old rat testis, named CG14, was hybridized in adult rat testicular section, in which the positive nucleic acid signals were distributed specifically in the primary spermatocytes. Another DNA fragment derived from 1 week old rat testis, named AA11, was hybridized specifically in Sertoli cell of 1 week old rat testis. Northern blot hybridization with [α 32 P] dCTP labeled CG14 probe, including cardiac, liver, kidney, brain, testis, and epididymis tissue mRNAs of rat, showed that an mRNA specific hybridization band, size of 1.258 kb, was found in testis tissue and size of 1.531 kb of another hybridization band present in epididymis tissue. The CG14 展开更多
基金supported by National Natural Science Foundation of China(No.3933140)Shanghai Municipal Education Commission(No.980802).
文摘AIM To find out the relationship between the gene transcription of different types ofprocollagen and the deposition of the relevant collagens in the liver tissue and to confirm the types of collagen producing cells in liverfibrogenesis.METHODS Dynamic changes of the expression of α1(Ⅰ ), α1 (Ⅲ ) and α1 (Ⅳ) procollagen mRNAand relevant collagens and the distribution ofcollagen producing cells during liver fibrogenesis of rat induced by CCl4 (20 weeks)were investigated with Northern blot analysis,in situ hybridization and immunohistochemicaltechniques.RESULTS The increased expression of α1 (Ⅲ)procollagen mRNA by Northern blot analysis was the most predominant one among the threemRNAs during fibrogenesis. However, theenhanced expression of al (Ⅳ) procollagenmRNA occurred very early while the expressionof α1 (Ⅰ) mRNA was not enhanced much until themiddle stage of the exPeriment. D6smin (Dm)positive hepatic stellate cells (HSCs) and fewmyofibroblasts (MFs) in and around the necrotic areas expressed α1 (Ⅰ), α1 (Ⅲ) and α1 (Ⅳ)procollagen mRNA signals detected by in situhybridization at the early stage of theexperiment. All the three procollagen mRNAsignals thereafter mainly localized in fibroblasts (Fbs) and MFs in fibrotic septa during the middle and late stages of fibrosis, which distributedparallel to the corresponding collagens detected by immunohistochemical study. ln addition, the endothelial cells of sinusoids and the small blood vessels within the septa also showed α1(Ⅳ) procollagen mRNA and type Ⅳ collagen expressionCONCLUSION It is considered that "HSC-MF-Fb" effect cell system is the major cellularsource of collagen production in liver fibrosis, in which HSCs are collagen producing precursor cells in the early liver fibrogenesis, thereafter the synthesis of type Ⅰ, Ⅲ and Ⅳ collagens (Col Ⅰ, Col Ⅲ and Col Ⅳ) mainly derives fromMFs and Fbs, which play a very important role in the progress of liver fibrosis. The endothelialcells along sinusoids, as another source of Col
文摘Kainic acid (KA) is a structural analogue of glutamate with potent excitotoxic andconvulsant properties. Intraperitoneal injection of KA in rats produces recurrentspontaneous limbic seizures accompanied by brain damage reminiscent of Ammon’s hornsclerosis, which has been considered as a useful model of human temporal lobe epilep-
文摘Objective To explore specific gene expression for regulating meiosis of germ cells during spermatogenesis of rat testis Materials & Methods Male SD rats, aged 1, 3 and 8 weeks, were observed in this study. The methods of morphological observation on testicular tissues embedded by resin and mRNA differential display (DDRT PCR) were combined to obtain specific mRNA expression gene fragments during the testicular development. Reverse dot blot hybridization was operated to further screen the positive differential DNA fragments. The positive DNA segments were sub cloned in pGEM T Easy vector and transformed into the competent E coli 109 straint. Northern blot analysis and in situ hybridization were also carried out for identifying tissue specific expression as well as cell specific expression DNA fragments. To screen λ ZAP II rat testicular gene library was searched for the original gene. Results Eighty two differential cDNA fragments were obtained through primary DDRT PCR, among which 40 differential cDNA fragments were selected for further screening with reverse dot blot hybridization. After the reverse dot blot hybridization, 12 primary differential DNA fragments were obtained. The size of DNA fragments ranged from 250 to 500 bp. The in situ hybridization of the testicular tissue showed that a specific DNA fragment derived from 8 week old rat testis, named CG14, was hybridized in adult rat testicular section, in which the positive nucleic acid signals were distributed specifically in the primary spermatocytes. Another DNA fragment derived from 1 week old rat testis, named AA11, was hybridized specifically in Sertoli cell of 1 week old rat testis. Northern blot hybridization with [α 32 P] dCTP labeled CG14 probe, including cardiac, liver, kidney, brain, testis, and epididymis tissue mRNAs of rat, showed that an mRNA specific hybridization band, size of 1.258 kb, was found in testis tissue and size of 1.531 kb of another hybridization band present in epididymis tissue. The CG14
