{"title":"大鼠脑切片在琼脂凝胶溶液中的长期可逆冷冻保存","authors":"A. Mokrushin","doi":"10.21638/spbu03.2023.103","DOIUrl":null,"url":null,"abstract":"Earlier there was found activity of glutamatergic ionotropic α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPAR) and N-methyl-D-aspartate receptors (NMDAR) disturbed after prolonged cryopreservation of brain slices at temperature of -10 оС. To eliminate cryodamage of AMPAR and NMDAR, the slices were encapsulated in the special freezing solution (SFS). SFS consisted of agar at various concentrations (33, 44 and 50 %) and artificial cerebrospinal fluid (67, 56 and 50 %, respectively). This solution was used for long-term cryopreservation of slices (52 days, -10 оС). Alterations in amplitudes of AMPA and NMDA potentials in the slices after rewarming were studied. Recovery of AMPAR and NMDAR after cryopreservation of the slices in SFS was the most optimal when the agar concentration in SFS was 50 %. Cryopreservation of the slices in SFS with different agar concentrations predominantly promoted the development of long-term potentiation in 78 % of the tested slices. Thus, the encapsulation of brain slices in SFS contributes to the preservation of AMPAR and NMDAR activity during long-term and reversible cryopreservation.","PeriodicalId":8998,"journal":{"name":"Biological Communications","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2023-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Encapsulation of rat brain slices in agar gel solution for long-term and reversible cryopreservation\",\"authors\":\"A. Mokrushin\",\"doi\":\"10.21638/spbu03.2023.103\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Earlier there was found activity of glutamatergic ionotropic α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPAR) and N-methyl-D-aspartate receptors (NMDAR) disturbed after prolonged cryopreservation of brain slices at temperature of -10 оС. To eliminate cryodamage of AMPAR and NMDAR, the slices were encapsulated in the special freezing solution (SFS). SFS consisted of agar at various concentrations (33, 44 and 50 %) and artificial cerebrospinal fluid (67, 56 and 50 %, respectively). This solution was used for long-term cryopreservation of slices (52 days, -10 оС). Alterations in amplitudes of AMPA and NMDA potentials in the slices after rewarming were studied. Recovery of AMPAR and NMDAR after cryopreservation of the slices in SFS was the most optimal when the agar concentration in SFS was 50 %. Cryopreservation of the slices in SFS with different agar concentrations predominantly promoted the development of long-term potentiation in 78 % of the tested slices. Thus, the encapsulation of brain slices in SFS contributes to the preservation of AMPAR and NMDAR activity during long-term and reversible cryopreservation.\",\"PeriodicalId\":8998,\"journal\":{\"name\":\"Biological Communications\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2023-05-02\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biological Communications\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.21638/spbu03.2023.103\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"Agricultural and Biological Sciences\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biological Communications","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.21638/spbu03.2023.103","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"Agricultural and Biological Sciences","Score":null,"Total":0}
引用次数: 0
摘要
早前发现,在-10 оС温度下长时间低温保存后,谷氨酸能离子型α-氨基-3-羟基-5-甲基异唑-4-丙酸(AMPAR)和n -甲基- d -天冬氨酸受体(NMDAR)的活性受到干扰。为了消除AMPAR和NMDAR的冷冻损伤,切片被包裹在特殊的冷冻溶液(SFS)中。SFS由不同浓度的琼脂(33%、44%和50%)和人工脑脊液(分别为67%、56%和50%)组成。该溶液用于切片的长期冷冻保存(52天,-10 оС)。研究了复温后各组AMPA和NMDA电位的变化。当琼脂浓度为50%时,在SFS中冷冻保存后AMPAR和NMDAR的回收率最高。在不同琼脂浓度的SFS中低温保存,78%的切片明显促进了长期增强的发展。因此,脑切片在SFS中的包封有助于AMPAR和NMDAR活性在长期和可逆冷冻保存期间的保存。
Encapsulation of rat brain slices in agar gel solution for long-term and reversible cryopreservation
Earlier there was found activity of glutamatergic ionotropic α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPAR) and N-methyl-D-aspartate receptors (NMDAR) disturbed after prolonged cryopreservation of brain slices at temperature of -10 оС. To eliminate cryodamage of AMPAR and NMDAR, the slices were encapsulated in the special freezing solution (SFS). SFS consisted of agar at various concentrations (33, 44 and 50 %) and artificial cerebrospinal fluid (67, 56 and 50 %, respectively). This solution was used for long-term cryopreservation of slices (52 days, -10 оС). Alterations in amplitudes of AMPA and NMDA potentials in the slices after rewarming were studied. Recovery of AMPAR and NMDAR after cryopreservation of the slices in SFS was the most optimal when the agar concentration in SFS was 50 %. Cryopreservation of the slices in SFS with different agar concentrations predominantly promoted the development of long-term potentiation in 78 % of the tested slices. Thus, the encapsulation of brain slices in SFS contributes to the preservation of AMPAR and NMDAR activity during long-term and reversible cryopreservation.