ÈÈÏߣº021-66110819,13564362870
Email£ºinfo@vizai.cn
ÈÈÏߣº021-66110819,13564362870
Email£ºinfo@vizai.cn
ÄÚÈÝÌáÒª£ºÄ¿µÄ£ºÑéÖ¤MED64ϵͳ¼Ç¼СÊóº£ÂíÄÔƬ×Ô·¢·Åµç·½·¨¿ÉÐÐÐÔ£¬ÒÔ¼°¹Û²ìƽÃæ΢µç¼«ÕóÁм¼ÊõÓ¦ÓÃÓÚÒÖÓô£¬ÅÁ½ðÉ£¬ñ²ðïµÈÑо¿µÄÓÅÊÆ¡£·½·¨£ºÍ¨¹ýMED64ϵͳ¶ÔÕý³£ÒÔ¼°ÓɵÍþ¸ß¼Ø¼Ó4-APÓÕµ¼µÄСÊóº£ÂíÄÔƬ½øÐмǼ£¬ÔËÓÃMobiusÈí¼þ½øÐÐÊý¾Ý·ÖÎö¡£½á¹û£ºÕý³£ACSF¹à×¢¼Ç¼µ½³ÉÄêСÊóº£ÂíÄÔƬÉñ¾Ôª×Ô·¢·Åµç¸ÄÓÃ4-AP+µÍþ¸ß¼ØACSF¹à×¢8min×óÓÒºóСÊóº£ÂíÄÔƬ³ÊÏÖñ²ðïÑù·Åµç¡£»»»ØÕý³£ACSFÏ´ÍÑ30minºóÄÔƬ·Åµç»Ö¸´Õý³££¬ñ²ðïÑù·ÅµçÖð½¥Ïûʧ¡£½áÂÛ£ºMED64ƽÃæ΢µç¼«ÕóÁм¼Êõ¿ÉÒԼǼСÊóº£ÂíÄÔƬÉñ¾Ôª×Ô·¢·Åµç£¬ÔËÓøü¼Êõ³É¹¦½¨Á¢ÁËСÊóº£ÂíÄÔƬñ²ðïÄ£ÐÍ¡£
΢µç¼«ÕóÁУ¨Multi-Electrode Array£©¼Ç¼ϵͳÊÇ°üÀ¨ÁËÀëÌåµçÉúÀíÑо¿ËùÐèµÄÒ»ÕûÌ×Ó²¼þÉ豸ºÄ²ÄºÍ·ÖÎöÈí¼þ¡£×Ô´ÓµÚÒ»Ì×΢µç¼«ÕóÁÐϵͳÎÊÊÀºó£¬ËüÖ÷ÒªÊÇͨ¹ýÈçºÎÈ¡µÃ¸üºÃµÄ°ûÍâ¼Ç¼ÐźźÍÔö¼Óµç¼«µÄÃܶÈÊýÁ¿ÉÏÕâÁ½·½ÃæÀ´·¢Õ¹µÄ¡£±¾Ð£ÖÐÐÄÊÔÑéÊÒµÄMED64ƽÃæ΢µç¼«ÕóÁÐÊÇÈÕ±¾Alpha MED Sciences¹«Ë¾Ñз¢µÄ¶àͨµÀÀëÌåµçÉúÀí¼Ç¼ϵͳ¡£Ëü¿Éͨ¹ý64¸öƽÃæ΢µç¼«¶ÔÀëÌå×éÖ¯»òϸ°û½øÐжàͨµÀµçÉúÀíͬ²½¼Ç¼¡£ÊÊÓÃÓÚÉñ¾¡¢ÊÓÍøĤºÍÐļ¡Ï¸°ûµçÉúÀíÌØÐÔºÍÀë×ÓͨµÀÉúÎïѧÌØÐÔÑо¿¡£ÓÉÓÚͬʱ¼Ç¼ÄÔƬ¶à¸öÉñ¾Ôª»ò¿ÉÐË·Ü×éÖ¯µÄ·Åµç£¬²É¼¯Êý¾Ý¶à£¬´ó´óµØÀ©Õ¹ÁËÑо¿µÄÊÓÒ°£¬ÓÐÖúÓÚ¶ÔÕû¸öÉñ¾ºËÍÅ»òÆ÷¹ÙµÄÉñ¾ÔªÖ®¼äÁªÏµºÍ¹¦ÄܵÄÑо¿¡£²¢ÇÒÓÉÓÚƽÃæ΢µç¼«ÕóÁм¼Êõ¶ÔÓÚϸ°ûÀ´ËµÊÇÎÞ´´ÐԵģ¬ÕâÑù¼È±£³ÖÁ˶Ե¥Ï¸°ûµÄ׼ȷ¼Ç¼£¬ÓÖÄܼǼ¶ÔÒ©Îï´Ì¼¤ÏìÓ¦µÄ½Ï³¤Ê±¼äµÄÑݱ䡣ÕâЩÌصãʹµÃÆä¾ßÓв»¿ÉÌæ´úµÄÓÅÊÆ¡£
Ä¿Ç°±¾Ð£ÒÑÓжà¸öÉñ¾ÉúÎïѧ¡¢ÉúÀíѧPI×é¶Ô¼Ç¼·ÖÎö»îÌå×éÖ¯Éñ¾Ôª×Ô·¢ÐԻÓÐÆÈÇÐÐèÇó£¬ÓÈÆäÊÇñ²ðﲡÑо¿ÁìÓò¡£Õâ¾ÍÐèÒª½¨Á¢Ò»Ì×ƽÃæ΢µç¼«ÕóÁмǼÀëÌåÉñ¾×éÖ¯×Ô·¢·ÅµçµÄ·½·¨¡£
1.²ÄÁÏÓë·½·¨
1.1Ò»°ã²ÄÁÏ
ʵÑ鶯ÎѡȡÇå½à¼¶½¡¿µ³ÉÄêÐÛÐÔC57BL/6СÊó£¬ÓÉÊ׶¼Ò½¿Æ´óѧ¶¯ÎﲿÌṩ£¬ÊµÑ鶯ÎïÐí¿ÉÖ¤ºÅ£ºSCXK£¨¾©£©2016-0011¡£ÊµÑ鶯ÎïµÄʹÓþù°´ÕÕÊ׶¼Ò½¿Æ´óѧʵÑ鶯ÎïÖÐÐĺͱ±¾©ÊµÑ鶯ÎïлáµÄ±ê×¼Ö´ÐС£Õû¸öʵÑé¹ý³ÌÖо¡¿ÉÄܵؼõÉÙ¶Ô¶¯ÎïµÄÉ˺¦¼°ËùÓö¯ÎïµÄÊýÁ¿¡£
ÊÔ¼Á£º4-°±»ùßÁण¨4-Aminopyridine£¬4-AP£©¹º×ÔSigma¹«Ë¾£¬ÆäÓàÊÔ¼Á¾ù¹º×Ô¹úÒ©¼¯ÍÅ»¯Ñ§ÊÔ¼ÁÓÐÏÞ¹«Ë¾¡£
1.2·½·¨
º£ÂíÄÔƬµÄÖƱ¸£º½«Ð¡Êó¶ÏÍ·£¬Ñ¸ËÙ°þÀë³öÈ«ÄÔÖÃÓÚ±ùË®»ìºÏ̬µÄ±ê×¼È˹¤ÄÔ¼¹Òº£¨Artificial Cerebrospinal Fluid£¬ACSF£©Öо²ÖÃ1min¡£ACSFµÄ×é³É£¨mmol/L£©£ºNaCl 117£¬KCl 3.6NaH2PO4¡¤2H2O 1.2£¬MgCl2¡¤6H2O 1.2£¬CaCl2¡¤2H2O2.5£¬NaHCO325£¬D-Glucose 11£¬pH 7.3¡«7.4¡£È»ºóÈ¡³öÊóÄÔ°þÀë³öº£Âí×éÖ¯ÖÁÓÚÐ޺õÄÇíÖ¬°¼²ÛÖУ¬ÓÃÂËÖ½Îü³ö¶àÓàÒºÌåºóÇеôºó¶ËС²¿×éÖ¯£¬¶øºóÓÃ502½º½«º£Âí×éÖ¯ÓëÇíÖ¬°¼²Û¹Ì¶¨ÓÚ×éÖ¯Õñ¶¯ÇÐƬ»ú£¨ÈÕ±¾Dosaka£¬DTK1000£©Ô¡²ÛµÄ½ðÊôµ×°åÉÏ£¬ÇÐÈ¡400¦ÌmºñµÄº£ÂíÄÔƬ£¨ÒÔ¹Ú×´ÃæÇÐƬ£©¡£½«ÇкõÄÄÔƬÒÆÖÁACSFµÄ·õÓý²ÛÖУ¬ACSFÔ¤ÏÈͨÒÔ95%O2ºÍ5%CO2»ìºÏÆøÖÁÑõ±¥ºÍ¡£ÊÒÎÂÏ·õÓý1hºó¼Ç¼¡£¼Ç¼ʱ³ÖÐø¹àÁ÷Ñõ±¥ºÍµÄACSF£¬Á÷ËÙ¿ØÖÆÔÚ3¡«4mL/min¡£
1.3 MED64ƽÃæ΢µç¼«ÕóÁеÄԤ׼±¸
±¾ÊµÑéµçÉúÀí¼Ç¼Ëù²ÉÓõÄMED64 Probe£¨MED64ƽÃæ΢µç¼«ÕóÁÐ̽Õ룬ÈÕ±¾Alpha Med Sciences£©ÊÇÔÚ͸Ã÷ʯӢ²£Á§£¨50mm¡Á50mm¡Á1.1mm£¬Pyrex7059£©ÖÐÐÄÄÚǶһ¸ö×°ÅäÓÐ64¸ö£¨8¡Á8µÈ¾àÕóÁУ©Æ½Ãæ΢µç¼«µÄµç·°å¹¹³É£¨Í¼1a£©[6]¡£MED64 ProbeÖÐÐÄÿ¸öƽÃæ΢µç¼«µÄÃæ»ýÊÇ50¦Ìm¡Á50¦Ìm£¬¼ä¾àΪ150¦Ìm£¨P515AÐͺţ©£¨Í¼1b£©¡£ÊµÑéÖиù¾Ý×éÖ¯±ê±¾´óС£¬Ãæ»ýµÄ²»Í¬¿ÉÒÔÑ¡ÔñÆäÊʺϵĵ缫¹æ¸ñÐͺš£µç¼«Ê¹ÓÃÇ°ÐèÒªÓú¬0.1%µÄ¾Ûµª±ûण¨Polyethylenimine£¬PEI£©Áò»¯ÎïµÄÅðËáÑλº³åÒº°üÂñ´¦Àí£¨12hÒÔÉÏ£©£¬Ä¿µÄÊǼÓÇ¿µç¼«±íÃæ¶ÔÄÔƬ×éÖ¯µÄð¤×ÅÇ׺ÍÐÔ¡£
ͼ1.MED64ƽÃæ΢µç¼«ÕóÁÐ̽Õ루a.MED64Probe£»b.8¡Á8µÈ¾à΢µç¼«ÕóÁУ»c.º£ÂíÄÔƬÔڵ缫ÕóÁÐÉϵÄλÖã©
1.4¼Ç¼º£ÂíÄÔƬ×Ô·¢·Åµç
Óÿí¿ÚÎü¹Ü½«·õÓýºÃµÄº£ÂíÄÔƬÒÆÈëMED64 Probe¡£ÔÚµ¹ÖÃÏÔ΢¾µÏÂÏÈÓÃÄáÁú¸ÇÍø¸ÇסÄÔƬ£¬È»ºóÓÃÄ÷×Ó¼ÐסÄáÁúÍø½«ÄÔƬÍÏÖÁµç¼«ÕóÁмǼλÖúó¸ÇÉϽðÊôѹƬ£¨SHD-22L£¬ÃÀ¹úHarvard£©½«Æä¹Ì¶¨¡£ÓÃÈ䶯±Ã£¨BT100-2J£¬LongPump£©³ÖÐø¹àÁ÷ͨÈë95%O2ºÍ5%CO2»ìºÏÆøµÄACSF£¨¹àÁ÷ËÙ¶È3¡«4mL/min£¬ÊÒΣ©¡£¹àÁ÷¹ý³ÌÖÐÓÃCCDÏÔ΢ÕÕÏà»ú£¨DP70£¬ÈÕ±¾Olympus£©²É¼¯º£ÂíÄÔƬ-MED64µç¼«Î»ÖÃͼÏñ£¨Í¼1c£©²¢ÇÒÅųýϵͳÔëÉù¸ÉÈÅ£¬½«ÔëÉù¿ØÖÆÔÚ¡À10¦ÌVÒÔÏ¡£¹àÁ÷20minºóʹÓÃMobiusÈí¼þ¶Ôº£ÂíÄÔƬÉñ¾ÔªµÄ×Ô·¢µç»î¶¯ÐźŽøÐÐͬ²½²ÉÑù£¬²ÉÑùÂÊΪ20kHz[7]¡£µÃµ½Õý³£×Ô·¢·ÅµçÊý¾Ýºó¸ÄÓÃ4-AP+µÍþ¸ß¼ØACSF¹àÁ÷£¬ÒÔÐγÉñ²ðïÑù·ÅµçµÄº£ÂíÄÔƬ£¨ñ²ðïÑù·Åµç´ïµ½10´Î/minÒÔÉÏ£©¡£¼Ç¼10minºó»»»ØÕý³£ACSFÏ´ÍÑÖ±ÖÁñ²ðïÑù·ÅµçÏûʧ¡£
2.½á¹û
2.1Õý³£Ð¡Êóº£ÂíÄÔƬ×Ô·¢·Åµç
Õý³£ACSF¹àÁ÷ʱ£¬MED64 Probe64¸öͨµÀÖнöÓиö±ðͨµÀ¿ÉÒԹ۲쵽º£ÂíÄÔƬÉÙÁ¿Éñ¾Ôª×Ô·¢·Åµç£¨Í¼2a-1£©¡£Í¨¹ýMobiusÈí¼þ¶Ô·ÅµçÐźÅͨµÀ·ÖÎö¿ÉÒÔÌáÈ¡³ö²»Í¬·ÅµçÌØÕ÷µÄ¶à¸öÉñ¾Ôª£¨Í¼2a-2£©¡£
ͼ2.СÊóº£ÂíÄÔƬ·Åµç£¨a-1.MED64¼Ç¼Õý³£Ð¡Êóº£ÂíÄÔƬ×Ô·¢·Åµç£»a-2.µÚ29ͨµÀº£ÂíÄÔƬÉñ¾Ôª×Ô·¢·Åµç£»b-1.MED64¼Ç¼4AP+ÎÞþ¸ß¼ØACSFÓÕµ¼µÄº£ÂíÄÔƬ×Ô·¢ÐÔñ²ðïÑù·Åµç£»b-2.µÚ27ͨµÀº£ÂíÄÔƬÉñ¾Ôªñ²ðïÑù·Åµç£©
2.2СÊóº£ÂíÄÔƬñ²ðïÑù·Åµç
¹àÁ÷4-AP+µÍþ¸ß¼ØACSF´óÔ¼8min×óÓÒ£¨£¨500¡À50£©s£¬n=6£©º£ÂíÄÔƬÉñ¾Ôª¿ªÊ¼³öÏÖ´Ø×´Ãܼ¯µÄñ²ðïÑù·Åµç£¨Í¼2b-1£©¡£ÔÚ³öÏÖÏà¶Ô´óµÄñ²ðïÑù·Åµçʼþ֮ǰ»áÏȹ۲쵽һЩͨµÀµÄÉñ¾ÔªÓɾ²Ä¬×ªÎªÐË·Ü£¬ÏȳöÏÖÒ»¶¨ÊýÁ¿µÄ×Ô·¢·Åµç£¬È»ºó³öÏÖÏà¶Ô·ù¶È½ÏС¼äÆڽϳ¤µÄñ²ðïÑù·Åµç¡£ñ²ðïÑù·ÅµçµÄÇøÓò°üÀ¨º£Âí£¨Cornu Ammonis£¬CA£©²¿·ÖÇøÓòºÍ³Ý×´»Ø£¨Dentate Gyrus£¬DG£©²¿·ÖÇøÓò£¬Éñ¾Ôª·Åµç·ùÖµÔÚ20¡«120¦ÌV£¨Í¼2b-2£©¡£¼Ç¼10minºó»»»ØÕý³£ACSFÏ´ÍÑ£¬ñ²ðïÑù·ÅµçËæÖ®Öð½¥¼õÈõ30minºóÍêÈ«Ïûʧ¡£
3.ÌÖÂÛ
ÏÖÔÚÉñ¾¿ÆѧÑо¿µÄÖ÷ҪĿ±ê¾ÍÊÇÑо¿¹¦ÄÜÐÔµÄÉñ¾»Ø·ÔÚÉúÀíºÍ²¡ÀíÇé¿öϵÄÇø±ð£¬¶øµ¥¸öÉñ¾ÔªÊǹ¹³ÉÉñ¾»Ø·µÄ»ù´¡¡£Éñ¾ÔªµÄ×Ô·¢µç»î¶¯ÊÇ´óÄÔÉñ¾»Ø·±àÂë»úÖƺÍÐÅÏ¢´¦Àí»úÖÆÑо¿µÄ»ù´¡£¬²úÉúÕâЩ×Ô·¢µç»î¶¯µÄÉúÀíÐźÅÊÇ·Ö²¼ÔÚ´óÄÔ¿Õ¼äµÄ£¬²¢ÇÒÍùÍù°´ÕÕijЩ¹æÔò·Ö²¼ÔÚ´óÄԵġ£Éñ¾ÏµÍ³¼²²¡µÄ·¢Éú·¢Õ¹ÍùÍù±íÏÖΪ×Ô·¢µç»î¶¯µÈÉúÀíÐźŵÄÒì³££¬ÔÚÉñ¾ÏµÍ³¼²²¡·¢ÉúµÄʱºòÍùÍù²»ÊÇÒ»¸öÉñ¾ÔªµÄ×Ô·¢µç»î¶¯·¢ÉúÁ˱仯£¬¶øÊǺܶàÉõÖÁÕû¸öºËÍŵÄ×Ô·¢µç»î¶¯ºÍ¹æÂÉ·¢ÉúÁ˱仯£¬±ÈÈç˵ÒÖÓô¡¢ñ²ðï¡¢ÅÁ½ðÉ¡¢°¢¶û´Äº£Ä¬µÈ¼²²¡¡£ËùÒÔͬʱ¶Ô×Ô·¢µç»î¶¯µÄ¿Õ¼ä·Ö²¼µÄ¶àͨµÀ¼Ç¼ÊǺÜÖØÒªµÄ¡£
Ä¿Ç°¶ÔÓÚÉñ¾Ôªµç»î¶¯µÄÑо¿·½·¨Ö÷ÒªÓÐϸ°ûÄÚºÍϸ°ûÍâ²£Á§µç¼«¼Ç¼£¬Î¢µç¼«ÕóÁм¼Êõ£¬¹âÒÅ´«Ñ§µÈ¡£Ï¸°ûÄÚºÍϸ°ûÍâ²£Á§µç¼«¼Ç¼ÊÇÄ¿Ç°×î³£Óõģ¬µ«Ö»ÄܼǼµ¥Ï¸°ûµÄ·Åµç£¬²»ÊÊÓÚÑо¿¶à¸öÉñ¾ÔªµÄµç»î¶¯£¬¶øÇÒËü¶ÔÓÚϸ°ûÀ´ÊÇÓд´µÄ£¬Ö»ÄÜÓÃÓÚ¶Ìʱ¼ä¼Ç¼¡£¹âÒÅ´«Ñ§Ö÷ÒªÓ¦ÓöԵçѹ»ò¸ÆÃô¸ÐµÄȾÁÏÀ´¹Û²ìϸ°ûµÄ»î¶¯£¬Ö»ÄÜ¿´ºÜdz²ãµÄÉñ¾Ôª£¨ÓÉÓÚÈëÉä¹âÖ»ÄÜ´©Í¸ºÜdzµÄÒ»²ã×éÖ¯£©£¬»¹ÓÐÐÔÔë±ÈµÄÎÊÌ⣬ÕâЩ¶¼ÏÞÖÆÁËËüµÄÓ¦Óá£ÁíÍ⻹ÓбÈÈ繦ÄÜÐԺ˴Ź²Õñ¡¢ÄÔµçͼ¡¢ÄÔƤ²ãµçͼ£¬µ«ÕâЩ·½·¨¿Õ¼ä·Ö±æÂʲ»¹»£¬¿´²»µ½µ¥¸öÉñ¾Ôªµç»î¶¯µÄ±ä»¯¡£Òª½ÒʾÉñ¾ÏµÍ³´¦ÀíÐÅÏ¢µÄ¸´ÔÓ»úÖÆ£¬±ØÐë»ñµÃ×ã¹»ÊýÄ¿µÄÉñ¾Ï¸°ûµÄµç»î¶¯ÐÅÏ¢¡£½üЩÄêËæ×ŵç×Ó¼¼ÊõºÍ΢»úе¼Ó¹¤¼¼ÊõµÄ¿ìËÙ·¢Õ¹£¬Î¢µç¼«ÕóÁм¼ÊõÒ²ËæÖ®µÃµ½Á˾޴óÌáÉý¡£Ä¿Ç°¸Ã¼¼Êõ¿ÉÒÔͬʱ¼ì²â¼¸Ê®ÉõÖÁÉÏ°Ù¸öͨµÀµÄµçÐźţ¬ÊµÏÖÁ˶ÔÉñ¾ÍøÂçµÄ¸ß¿Õ¼ä·Ö±æÂʼì²â¡£Òò´Ë£¬MED64ƽÃæ΢µç¼«ÕóÁÐϵͳ¶ÔÑо¿Éñ¾×éÖ¯ÍøÂçÌØÐÔÓÐןܴóµÄ¼¼ÊõÓÅÊÆ¡£
΢µç¼«ÕóÁм¼Êõ½üÄêÀ´¶àÓÃÓÚÅàÑøÉñ¾Ôª¼°Éñ¾×éÖ¯µÄµçÉúÀí¼Ç¼¡£¼±ÐÔÄÔƬµÄ¼Ç¼Ҳ¶àΪ¼Ç¼ÐË·ÜÐÔÍ»´¥ºó³¡µç루Field Excitatory Postsynaptic Potential£¬fEPSP£©¡£¼±ÐÔÄÔƬ×Ô·¢·Åµç¼Ç¼ÉÙÓб¨µÀ¡£±¾ÊµÑéʹÓÃƽÃæ΢µç¼«ÕóÁм¼Êõ¼Ç¼µ½³ÉÄêСÊóº£ÂíÄÔƬÉñ¾Ôª×Ô·¢·Åµç²¢Çҳɹ¦½¨Á¢4-AP+µÍþ¸ß¼ØACSFÓÕµ¼µÄ¼±ÐÔСÊóº£ÂíÄÔƬñ²ðïÄ£ÐÍ¡£Í¨¹ýʵÑéÍêÉÆÓÅ»¯ÁËMED64ϵͳ¼Ç¼ÀëÌåÄÔƬ×Ô·¢·ÅµçµÄ·½·¨£¬ÍØÕ¹Á˸ÃÉ豸¹¦ÄÜ¡£Îª¼Ç¼·ÖÎö»îÌå×éÖ¯Éñ¾Ôª×Ô·¢ÐԷŵç»î¶¯µÄÑо¿ÌṩÏàÓ¦µÄ¼¼Êõ±£ÕÏ¡£ÎªÑ§Ð£Ñо¿ñ²ðï¡¢ÅÁ½ðÉ¡¢ÒÖÓô¡¢°¢¶û´Äº£Ä¬²¡µÈÖØ´óÄÔ¼²²¡Ìṩ¿ÆѧÑо¿Æ½Ì¨¡£