Morpho-functional characterization of neuronal cells at different stages of maturation in granule cell layer of adult rat dentate gyrus.
Neurogenesis occurs throughout adult life in dentate gyrus of mammal hippocampus. Therefore, neurons at different stages of electrophysiological and morphological maturation and showing various, if any, synaptic inputs co-exist in the adult granule cell layer, as occurs during dentate gyrus development. The knowledge of functional properties of new neurons throughout their maturation can contribute to understanding their role in the hippocampal function. In this study electrophysiological and morphological features of granule layer cells, characterized as immature or mature neurons, without and with synaptic input, were comparatively described in adult rats. The patch-clamp technique was used to perform electrophysiological recordings, the occurrence of synaptic input evoked by medial perforant pathway stimulation was investigated and synaptic input was characterized. Cells were then identified and morphologically described via detection of biocytin injected through the patch pipette. The neuronal phenotype of recorded cells was assessed by immunohistochemistry and single-cell RT-PCR. Cells with very low capacitance, high input resistance, depolarized resting membrane potential and without synaptic activity were found exclusively at the border of the GCL facing hilus; this type of cell expressed the class III beta-tubulin neuronal marker (mRNA and protein) and did not express a glial marker. Immature neuronal cells with progressively increasing capacitance, decreasing input resistance and resting membrane potential getting more hyperpolarized showed only depolarizing GABAergic synaptic input at first and then also glutamatergic synaptic input. Finally, cells showing electrophysiological, synaptic, and morphological features of mature granule, expressing the mature neuron marker NeuN, were identified.
Ambrogini
Patrizia
P
Istituto di Scienze Fisiologiche, Università degli Studi di Urbino, Carlo Bo, Crocicchia, I-61029 Urbino, Italy. p.ambrogini@uniurb.it
Lattanzi
Davide
D
Ciuffoli
Stefano
S
Agostini
Deborah
D
Bertini
Luana
L
Stocchi
Vilberto
V
Santi
Spartaco
S
Cuppini
Riccardo
R
D018750:6-Cyano-7-nitroquinoxaline-2,3-dione / Q000494:pharmacology; D000818:Animals; D001640:Bicuculline / Q000031:analogs & derivatives* / Q000494:pharmacology; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002232:Carbocyanines; D016922:Cellular Senescence / Q000187:drug effects / Q000502:physiology*; D018891:Dentate Gyrus / Q000166:cytology*; D004558:Electric Stimulation / Q000379:methods; D018690:Excitatory Amino Acid Agonists / Q000494:pharmacology; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D018756:GABA Antagonists / Q000494:pharmacology; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D007150:Immunohistochemistry / Q000379:methods; D066298:In Vitro Techniques; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D018613:Microscopy, Confocal / Q000379:methods; D039842:Neural Cell Adhesion Molecule L1 / Q000378:metabolism; D009474:Neurons / Q000145:classification / Q000502:physiology*; D018408:Patch-Clamp Techniques / Q000379:methods; D019580:Perforant Pathway / Q000528:radiation effects; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012794:Sialic Acids / Q000378:metabolism; D013569:Synapses / Q000187:drug effects / Q000502:physiology / Q000528:radiation effects; D009435:Synaptic Transmission / Q000187:drug effects / Q000502:physiology*; D013779:Tetrodotoxin / Q000494:pharmacology; D014404:Tubulin / Q000235:genetics / Q000378:metabolism; D014633:Valine / Q000031:analogs & derivatives* / Q000494:pharmacology; D014700:Verapamil / Q000494:pharmacology
D002121:Calcium Channel Blockers; D002232:Carbocyanines; D018690:Excitatory Amino Acid Agonists; D018691:Excitatory Amino Acid Antagonists; D018756:GABA Antagonists; D005904:Glial Fibrillary Acidic Protein; D039842:Neural Cell Adhesion Molecule L1; D012333:RNA, Messenger; D012794:Sialic Acids; D014404:Tubulin; C089187:cyanine dye 3; C103469:polysialyl neural cell adhesion molecule; C017069:bicuculline methiodide; D013779:Tetrodotoxin; D018750:6-Cyano-7-nitroquinoxaline-2,3-dione; C056639:2-amino-5-phosphopentanoic acid; D014700:Verapamil; D010751:Phosphopyruvate Hydratase; D014633:Valine; D001640:Bicuculline