A cDNA encoding the mouse GABA transporter has been isolated and sequenced. The results show that the mouse GABA transporter cDNA differs from that of the rat by 60 base pairs at the open reading frame region but the ...A cDNA encoding the mouse GABA transporter has been isolated and sequenced. The results show that the mouse GABA transporter cDNA differs from that of the rat by 60 base pairs at the open reading frame region but the deduced amino acid sequences of the two cDNAs are identical and both composed of 599 amino acids. However,the amino acid sequence is different from the sequence deduced from a recently published mouse GABA transporter cDNA.展开更多
The cDNA molecule encoding the mouse GABA transporter gene (GAT-1) was used as probe for selecting GAT-1 gene from mouse genomic library. A positive clone, harboring the whole open reading frame of the GAT-1 protein a...The cDNA molecule encoding the mouse GABA transporter gene (GAT-1) was used as probe for selecting GAT-1 gene from mouse genomic library. A positive clone, harboring the whole open reading frame of the GAT-1 protein and designated as MGABAT-G, was fished out from the library, the 5’ proximal region and nitron 1 were sequenced and analysed, and low homology was found in the above region between GAT-1 genes from mouse and human except some short conserved sequences. The DNA-protein interactions between DNA fragments containing the conserved sequences in the 5’ proximal region and nuclear proteins from different tissues of mouse were studied by means of gel-shift assay, and Southern-Western blot. The results indicate a possible positive-negative regulation mode controlling the expression of the mouse GAT-1 gene.展开更多
Objective To evaluate the effect of neuropathic pain on the expression of GABAB receptor subtypes in spinal cord of rats with chronic constriction injury.Methods 36 rats were divided into Sham group and CCI group resp...Objective To evaluate the effect of neuropathic pain on the expression of GABAB receptor subtypes in spinal cord of rats with chronic constriction injury.Methods 36 rats were divided into Sham group and CCI group respectively.In CCI group,the left common sciatic nerve of anesthetized rats was exposed,four silk ligatures（30） were tied loosely proximal to the sciatica’s trifurcation.In Sham group,Sham surgery was undergone by exposing the left sciatic nerve without ligation.Hind paw thermal withdrawl latency（TWL） to thermal stimulus and mechanical withdrawl threshold（MWT） of mechanical stimulus were recorded before ligation or on 1 d,3 d,5 d,7 d,10 d,14 d,21 d,28 d after ligation respectively（n=8）.Expression of GABAB receptor subtypes were detected before ligation and 1 d,3 d,7 d,14 d,28 d after ligation respectively using bothe immunohistochemical assay（n=6） and Western blot technique（n=4）.Results There were no significant differences on the values of MWT and TWL before ligation between Sham group and CCI group.Compared with Sham group,the MWT and TWL of rats in CCI group significantly decreased after ligation（P<0.01,P<0.05） and continued to be decreased until 28 d after ligation.Both immunoreactive neurons and protein content of GABAB1a or GABAB2 subtypes increased on the 3rd day after ligation,then both decreased on the 7th day after ligation,and lasted for 28 dafter ligation.（P<0.01,P<0.05）.There were no changes of GABAB1b after ligation（P>0.05）.Conclusion There existed differences among expression of spinal GABAB receptor subtypes on rats with chronic constriction injury induced neuropathic pain.The expression of spinal GABAB1a and GABAB2 decreased while there were no changes of GABAB1b on neuropathic rats.展开更多
Transgenic mice ubiquitously overexpressing murine γ aminobutyric acid transporter subtype Ⅰ were created. Unexpectedly, these mice markedly exhibited heritable obesity, which features significantly increased body w...Transgenic mice ubiquitously overexpressing murine γ aminobutyric acid transporter subtype Ⅰ were created. Unexpectedly, these mice markedly exhibited heritable obesity, which features significantly increased body weight and fat deposition. Behavioral examination revealed that transgeinc mice have slightly reduced spontaneous locomotive capacity and altered feeding pattern. Tills preliminary finding indicates that the inappropriate level of γ-aminobutyric acid transporters may be directly or indirectly involved in the pathogenic mechanism underlying certain types of obesity.展开更多
Some recent studies indicated that GABAergic system is involved in mammalian sperm acrosome reaction (AR), but direct evidence pertaining to the expression of gat1 in mammalian sperm is not yet demonstrated. In this s...Some recent studies indicated that GABAergic system is involved in mammalian sperm acrosome reaction (AR), but direct evidence pertaining to the expression of gat1 in mammalian sperm is not yet demonstrated. In this study, we evaluated the presence of 67kDa GAT1 protein and mRNA in rat testis by Western blotting and reverse transcription-polymerase chain reaction. Meanwhile, immunohistochemical and immunofluorescent analyses also identified GAT1 protein on the elongated spermatid and sperm. These results indicated that rat testis is a novel site of gat1 expression. Further studies should be taken to explore the role of GAT1 protein on sperm acrosome reaction.展开更多
γ-Aminobutyric acid and GABAergic receptors were previously reported to be distributed in reproductive systems besides CNS and predicted to participate in the modulation of testicular function. γ-Aminobutyric acid t...γ-Aminobutyric acid and GABAergic receptors were previously reported to be distributed in reproductive systems besides CNS and predicted to participate in the modulation of testicular function. γ-Aminobutyric acid transporter was implicated to be involved in this process. However, the potential role of γ-aminobutyric transporter in testis has not been explored. In this study, we investigated the existence of mouse γ-aminobutyric acid transporter subtype I (mGAT1) in testis. Wild-type and transgenic mice, which overexpressing mGAT1 in a variety of tissues, especially in testis, were primarily studied to approach the profile of mGAT1 in testis. Mice with overexpressed mGAT1 develop normally but with reduced mass and size of testis as compared with wild-type. Testicular morphology of transgenic mice exhibited overt abnormalities including focal damage of the spermatogenic epithelium accompanied by capillaries proliferation and increased diameter of seminiferous tubules lumen. Reduced number of spermatids was also found in some seminiferous tubules. Our results clearly demonstrate the presence of GAT1 in mouse testis and imply that GAT1 is possibly involved in testicular function.展开更多
文摘A cDNA encoding the mouse GABA transporter has been isolated and sequenced. The results show that the mouse GABA transporter cDNA differs from that of the rat by 60 base pairs at the open reading frame region but the deduced amino acid sequences of the two cDNAs are identical and both composed of 599 amino acids. However,the amino acid sequence is different from the sequence deduced from a recently published mouse GABA transporter cDNA.
文摘The cDNA molecule encoding the mouse GABA transporter gene (GAT-1) was used as probe for selecting GAT-1 gene from mouse genomic library. A positive clone, harboring the whole open reading frame of the GAT-1 protein and designated as MGABAT-G, was fished out from the library, the 5’ proximal region and nitron 1 were sequenced and analysed, and low homology was found in the above region between GAT-1 genes from mouse and human except some short conserved sequences. The DNA-protein interactions between DNA fragments containing the conserved sequences in the 5’ proximal region and nuclear proteins from different tissues of mouse were studied by means of gel-shift assay, and Southern-Western blot. The results indicate a possible positive-negative regulation mode controlling the expression of the mouse GAT-1 gene.
文摘Objective To evaluate the effect of neuropathic pain on the expression of GABAB receptor subtypes in spinal cord of rats with chronic constriction injury.Methods 36 rats were divided into Sham group and CCI group respectively.In CCI group,the left common sciatic nerve of anesthetized rats was exposed,four silk ligatures（30） were tied loosely proximal to the sciatica’s trifurcation.In Sham group,Sham surgery was undergone by exposing the left sciatic nerve without ligation.Hind paw thermal withdrawl latency（TWL） to thermal stimulus and mechanical withdrawl threshold（MWT） of mechanical stimulus were recorded before ligation or on 1 d,3 d,5 d,7 d,10 d,14 d,21 d,28 d after ligation respectively（n=8）.Expression of GABAB receptor subtypes were detected before ligation and 1 d,3 d,7 d,14 d,28 d after ligation respectively using bothe immunohistochemical assay（n=6） and Western blot technique（n=4）.Results There were no significant differences on the values of MWT and TWL before ligation between Sham group and CCI group.Compared with Sham group,the MWT and TWL of rats in CCI group significantly decreased after ligation（P<0.01,P<0.05） and continued to be decreased until 28 d after ligation.Both immunoreactive neurons and protein content of GABAB1a or GABAB2 subtypes increased on the 3rd day after ligation,then both decreased on the 7th day after ligation,and lasted for 28 dafter ligation.（P<0.01,P<0.05）.There were no changes of GABAB1b after ligation（P>0.05）.Conclusion There existed differences among expression of spinal GABAB receptor subtypes on rats with chronic constriction injury induced neuropathic pain.The expression of spinal GABAB1a and GABAB2 decreased while there were no changes of GABAB1b on neuropathic rats.
文摘Transgenic mice ubiquitously overexpressing murine γ aminobutyric acid transporter subtype Ⅰ were created. Unexpectedly, these mice markedly exhibited heritable obesity, which features significantly increased body weight and fat deposition. Behavioral examination revealed that transgeinc mice have slightly reduced spontaneous locomotive capacity and altered feeding pattern. Tills preliminary finding indicates that the inappropriate level of γ-aminobutyric acid transporters may be directly or indirectly involved in the pathogenic mechanism underlying certain types of obesity.
基金grants of National "Pan Deng" program LMCBand National Natural Science FOundation!No: 39770776
文摘Some recent studies indicated that GABAergic system is involved in mammalian sperm acrosome reaction (AR), but direct evidence pertaining to the expression of gat1 in mammalian sperm is not yet demonstrated. In this study, we evaluated the presence of 67kDa GAT1 protein and mRNA in rat testis by Western blotting and reverse transcription-polymerase chain reaction. Meanwhile, immunohistochemical and immunofluorescent analyses also identified GAT1 protein on the elongated spermatid and sperm. These results indicated that rat testis is a novel site of gat1 expression. Further studies should be taken to explore the role of GAT1 protein on sperm acrosome reaction.
基金grants from National Science Foundation!No.39630140
文摘γ-Aminobutyric acid and GABAergic receptors were previously reported to be distributed in reproductive systems besides CNS and predicted to participate in the modulation of testicular function. γ-Aminobutyric acid transporter was implicated to be involved in this process. However, the potential role of γ-aminobutyric transporter in testis has not been explored. In this study, we investigated the existence of mouse γ-aminobutyric acid transporter subtype I (mGAT1) in testis. Wild-type and transgenic mice, which overexpressing mGAT1 in a variety of tissues, especially in testis, were primarily studied to approach the profile of mGAT1 in testis. Mice with overexpressed mGAT1 develop normally but with reduced mass and size of testis as compared with wild-type. Testicular morphology of transgenic mice exhibited overt abnormalities including focal damage of the spermatogenic epithelium accompanied by capillaries proliferation and increased diameter of seminiferous tubules lumen. Reduced number of spermatids was also found in some seminiferous tubules. Our results clearly demonstrate the presence of GAT1 in mouse testis and imply that GAT1 is possibly involved in testicular function.
