Bernstein Hannah L, Lu Yi-Ling, Botterill Justin J, Duffy Áine M, LaFrancois John J, Scharfman Helen E
bioRxiv. 2024 Nov 8:2024.11.08.622679. doi: 10.1101/2024.11.08.622679.
Glutamatergic dentate gyrus (DG) mossy cells (MCs) innervate the primary DG cell type, granule cells (GCs). Numerous MC synapses are on GC proximal dendrites in the inner molecular layer (IML). However, field recordings of the GC excitatory postsynaptic potential (fEPSPs) have not been used to study this pathway selectively. Here we describe methods to selectively activate MC axons in the IML using mice with Cre recombinase expressed in MCs. Slices were made after injecting adeno-associated virus (AAV) encoding channelrhodopsin (ChR2) in the DG. In these slices, we show that fEPSPs could be recorded reliably in the IML in response to optogenetic stimulation of MC axons. Furthermore, fEPSPs were widespread across the septotemporal axis. However, fEPSPs were relatively weak because they were small in amplitude and did not elicit a significant population spike in GCs. They also showed little paired pulse facilitation. We confirmed the extracellular findings with patch clamp recordings of GCs despite different recording chambers and other differences in methods. Together the results provide a simple method for studying MC activation of GCs and add to the evidence that this input is normally weak but widespread across the GC population.
We describe a method to activate the MC input to GCs selectively using optogenetics in hippocampal slicesMC excitation is weakly excitatory but so common among GCs that a field EPSP is generated at the site of MC synapses on GCsMC excitation of GCs is consistent across the septotemporal axis and contralaterallyUsing the characteristics of optogenetically-evoked fEPSPs, electrical stimulation of the MC input to GCs can be optimized.
谷氨酸能齿状回(DG)苔藓细胞(MCs)支配主要的DG细胞类型,即颗粒细胞(GCs)。许多MC突触位于内分子层(IML)的GC近端树突上。然而,GC兴奋性突触后电位(fEPSPs)的场记录尚未被用于选择性地研究这条通路。在此,我们描述了使用在MCs中表达Cre重组酶的小鼠选择性激活IML中MC轴突的方法。在DG中注射编码通道视紫红质(ChR2)的腺相关病毒(AAV)后制作切片。在这些切片中,我们表明,响应于MC轴突的光遗传学刺激,可以在IML中可靠地记录fEPSPs。此外,fEPSPs在颞-中隔轴上广泛分布。然而,fEPSPs相对较弱,因为它们的幅度较小,并且在GCs中不会引发明显的群体峰电位。它们也几乎没有成对脉冲易化现象。尽管记录室不同且方法存在其他差异,我们通过GCs的膜片钳记录证实了细胞外实验结果。这些结果共同提供了一种研究MCs对GCs激活作用的简单方法,并进一步证明了这种输入通常较弱,但在整个GC群体中广泛存在。
我们描述了一种在海马切片中使用光遗传学选择性激活MCs对GCs输入的方法
MCs兴奋是弱兴奋性的,但在GCs中非常普遍,以至于在GCs上MC突触部位产生场兴奋性突触后电位
MCs对GCs的兴奋在颞-中隔轴上和对侧是一致的
利用光遗传学诱发的fEPSPs的特征,可以优化对MCs输入到GCs的电刺激。