Morphologic and electrophysiologic maturation in developing dentate gyrus granule cells.

Dentate gyrus granule cells from immature (7-28 days) Sprague-Dawley rats were examined with whole cell patch clamp recordings and biocytin filling in in vitro hippocampal slice preparations. Although recordings were confined to the middle third of the suprapyramidal limb of the dentate, the granule cells exhibited marked variability in their physiologic properties: input resistance (IR) ranged from 250 MOmega to 3 GOmega, and resting membrane potential (RMP) from -82 to -41 mV. Both IR and RMP were inversely correlated with dendritic length, a morphometric indicator of cell maturity. Thus the highest IR cells were the youngest, and maturation was characterized by a progressive decrease in IR, hyperpolarization of RMP, and elongation of the dendritic arbor. When cells were grouped by IR, significant intergroup differences were found in RMP, dendritic length, and number of dendritic terminal branches. Although cells of all IR categories were examined throughout the age spectrum under study, none of the inter-IR group differences was age-dependent. These data suggest that IR provides a reasonable estimate of granule cell maturity and that maturation entails predictable changes in cell properties and morphology. These aspects of maturation correlate with each other, are independent of animal age, and most likely proceed according to a program related to cell birth.

Developmental changes in membrane properties and postsynaptic currents of granule cells in rat dentate gyrus.

1. Whole cell patch-clamp recordings were used to study dentate gyrus granule cells in hippocampal slices from juvenile rats (postnatal days 8-32). Membrane properties were measured with the use of current-clamp recordings and were correlated with the morphology of a subgroup of neurons filled with biocytin. The components of the postsynaptic currents (PSCs) induced by medial perforant path stimulation were characterized with the use of specific receptor antagonists in voltage-clamp recordings. 2. Granule cells located in the middle third of the superior blade of stratum granulosum from the rostral third of hippocampus were divided into three groups according to their input resistance (IR). Neurons with low IR (206 +/- 182 M omega, mean +/- SD) had hyperpolarized resting membrane potentials (-82 +/- 7 mV) and high-amplitude action potentials (108 +/- 23 mV). Neurons were high IR (1,259 +/- 204 M omega) had more depolarized resting membrane potentials (-54 +/- 6 mV) and lower-amplitude action potentials (71 +/- 10 mV). Neurons with intermediate IR (619 +/- 166 M omega) also had intermediate resting membrane potentials (-63 +/- 7 mV) and action potential amplitudes (86 +/- 14 mV). Low-IR neurons became increasingly prevalent with advancing postnatal age, but neurons from each group could be found throughout the entire period under study. 3. Morphological studies of low-IR neurons revealed an extensive dendritic arborization that traversed the entire molecular layer and was characteristic of mature granule cells. High-IR cells had smaller somata and short, simple dendritic arborization that incompletely penetrated the molecular layer and were classified as immature. Intermediate-IR cells had morphological features of intermediate maturity. 4. The initial phase of the PSC evoked at -80 mV was a fast inward current that was comparable with respect to latency to peak, latency to onset, and 10-90% rise time in neurons of all maturities held at -80 mV. This current was 6-cyano-7-nitroquinoxaline-2,3-dione sensitive. 5. The decay phases of PSCs at -80 mV varied with neuronal maturity. Mature neurons had monoexponential decays (tau = 8.9 +/- 3.6). Intermediate and immature neurons had prominent later inward currents that resulted in slower decays. In the case of the immature neurons, the inward current during the decay phase could be separated from the initial fast inward peak. The later inward currents in intermediate and immature neurons were bicuculline sensitive. 6. With the use of uniform ionic conditions of the extracellular and patch solutions, current-voltage relations and reversal potentials for pharmacologically isolated alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), N-methyl-D-aspartate (NMDA), and gamma-aminobutyric acid-A (GABAA) currents were comparable across all cell maturities. Calculated ratios for peak GABAA/NMDA/AMPA currents decreased significantly with maturation as follows: 9.4 +/- 2.9/1.4 +/- 0.5/1.0 for immature cells, 7.2 +/- 2.5/1.5 +/- 0.7O/1.0 for intermediate cells, and 2.0 +/- 1.2/0.9 +/- 0.4/1.0 for mature cells. 7. GABA current was mediated both by polysynaptic activation of interneurons and by direct activation of interneurons with monosynaptic input onto granule cells. The proportional contributions of mono- and polysynaptic GABA to total GABA were comparable across all cell maturities; latency to peak GABA current decreased with increasing cell maturity for both mono- and polysynaptic components. 8. We conclude that PSCs evoked in granule cells by medial perforant path activation in neurons of all maturities consist of both glutamatergic and GABAergic components. PSCS are dominated by GABAergic neurotransmission in immature granule cells, and the contribution of glutamatergic neurotransmission increases with neuronal maturation. The greater ratio of peak GABAA to glutamate currents and the longer time interval between their respective peaks combine to produce a distinctive PSC shape