Electrophysiological Properties of Cultured Neonatal Rat Dorsal Horn Neurons Containing GABA and Met-Enkephalin-Like Immunoreactivity

1998 ◽  
Vol 79 (3) ◽  
pp. 1583-1586 ◽  
Author(s):  
Y. H. Jo ◽  
M. E. Stoeckel ◽  
R. Schlichter

Jo, Y. H., M. E. Stoeckel, and R. Schlichter. Electrophysiological properties of cultured neonatal rat dorsal horn neurons containing GABA and met-enkephalin-like immunoreactivity. J. Neurophysiol. 79: 1583–1586, 1998. We have developed a culture of neurons dissociated from the most superficial laminae of the neonatal rat spinal cord dorsal horn (DH). By using the perforated patch-clamp technique, we distinguished four types of neurons based on their firing properties in response to intracellular injection of 900 ms lasting current pulses. Type 1 neurons were characterized by a tonic firing. Type 2 neurons displayed marked spike accommodation and fired brief (<500 ms) bursts of action potentials, whereas type 3 neurons fired a single spike. Type 4 neurons exhibited different types of firing patterns, but all of them possessed a time-dependent inwardly rectifying current activated by membrane hyperpolarization. Met-enkephalin-like immunoreactivity (met-ENK-LI) and glutamic acid decarboxylase-like immunoreactivity (GAD-LI) were colocalized in 42% of the neurons ( n = 59), which were previously identified electrophysiologically. Type 1–4 neurons represented respectively 4, 64, 20, and 12% of the population of neurons colocalizing met-ENK-LI and GAD-LI. We conclude that the electrophysiological properties of DH neurons present in our cultures are similar to those described in acute slice or hemisected spinal cord preparations and that met-ENK-LI and GABA-LI are preferentially colocalized in type 2 neurons.

1996 ◽  
Vol 76 (3) ◽  
pp. 1465-1476 ◽  
Author(s):  
B. A. Miller ◽  
C. J. Woolf

1. The role of glutamate in slow excitatory synaptic transmission between small-diameter primary afferents and deep dorsal horn neurons was examined in neonatal rat spinal cord in vitro with the use of the whole cell voltage-clamp technique. 2. Single-shock electrical stimulation of large-diameter A beta-fibers evoked a short-latency (< 10 ms) fast (< 500 ms) excitatory postsynaptic current (EPSC). Stimulation of small-diameter A delta- and C fibers resulted, in addition, in a slowly rising and decaying EPSC (lasting up to 14 s) following the fast EPSC. The slow EPSC was never observed with stimulation of A beta-fibers. 3. Two patterns of EPSCs were observed, "type 1" and "type 2," which differed in their time course (lasting up to 1 and 14 s, respectively). The type 1 response was biphasic, with a fast monosynaptic component followed by an invariant, presumably monosynaptic, late slow component. The type 2 response was multiphasic, with a fast monosynaptic component followed by a slow component composed of fast polysynaptic currents superimposed on a slow current. 4. The fast monosynaptic component had a linear conductance, whereas the late slower component of the A beta-fiber-evoked response had a negative slope conductance at holding potentials more negative than -23 mV. Both currents reversed at a membrane potential of -1.2 +/- 2.8 (SE) mV. 5. With the use of selective non-N-methyl-D-aspartate (non-NMDA) and NMDA receptor antagonists [6-cyano-7-nitroquinox-aline-2,3-dione (CNQX) or 2,3-dihydroxy-6-nitro-7-sulphamoyl-benzo (F) quinoxaline and D(-)-2-amino-5-phosphonopentanoic acid (D-AP5), respectively] we showed that both the early fast (A beta-fiber evoked) and the late slow (A delta- and C fiber evoked) components were mediated by non-NMDA and NMDA receptors. CNQX suppressed both the early fast and late slow components of the compound EPSC, whereas D-AP5 suppressed the polysynaptic currents of the early fast component and the late slow component without significantly affecting the early fast monosynaptic component. 6. Slow EPSCs summated on low-frequency (1 or 10 Hz), repetitive stimulation and produced long-duration "tail" currents on cessation of the stimulus. The amount of temporal summation was proportional to the duration of the slow EPSC and the frequency of stimulation. 7. Our results suggest that slow ionotropic-glutamate-receptor-mediated EPSCs produced by the stimulation of small-diameter primary afferents play an important role in activity-dependent synaptic plasticity in the dorsal horn.


2008 ◽  
Vol 99 (6) ◽  
pp. 3144-3150 ◽  
Author(s):  
Rachel A. Ingram ◽  
Maria Fitzgerald ◽  
Mark L. Baccei

The lower thresholds and increased excitability of dorsal horn neurons in the neonatal rat suggest that inhibitory processing is less efficient in the immature spinal cord. This is unlikely to be explained by an absence of functional GABAergic inhibition because antagonism of γ-aminobutyric acid (GABA) type A receptors augments neuronal firing in vivo from the first days of life. However, it is possible that more subtle deficits in GABAergic signaling exist in the neonate, such as decreased reliability of transmission or greater depression during repetitive stimulation, both of which could influence the relative excitability of the immature spinal cord. To address this issue we examined monosynaptic GABAergic inputs onto superficial dorsal horn neurons using whole cell patch-clamp recordings made in spinal cord slices at a range of postnatal ages (P3, P10, and P21). The amplitudes of evoked inhibitory postsynaptic currents (IPSCs) were significantly lower and showed greater variability in younger animals, suggesting a lower fidelity of GABAergic signaling at early postnatal ages. Paired-pulse ratios were similar throughout the postnatal period, whereas trains of stimuli (1, 5, 10, and 20 Hz) revealed frequency-dependent short-term depression (STD) of IPSCs at all ages. Although the magnitude of STD did not differ between ages, the recovery from depression was significantly slower at immature GABAergic synapses. These properties may affect the integration of synaptic inputs within developing superficial dorsal horn neurons and thus contribute to their larger receptive fields and enhanced afterdischarge.


2010 ◽  
Vol 6 ◽  
pp. 1744-8069-6-25 ◽  
Author(s):  
Stefan K Georgiev ◽  
Hidemasa Furue ◽  
Hiroshi Baba ◽  
Tatsuro Kohno

2001 ◽  
Vol 85 (5) ◽  
pp. 1864-1872 ◽  
Author(s):  
Sandra M. Garraway ◽  
Shawn Hochman

5-hydroxytryptamine (5-HT) is released in spinal cord by descending systems that modulate somatosensory transmission and can potently depress primary afferent-evoked synaptic responses in dorsal horn neurons. Since primary afferent activity-induced long-term potentiation (LTP) may contribute to central sensitization of nociception, we studied the effects of 5-HT on the expression of sensory-evoked LTP and long-term depression (LTD) in deep dorsal horn (DDH) neurons. Whole cell, predominantly current clamp, recordings were obtained from DDH neurons in transverse slices of neonatal rat lumbar spinal cord. The effect of 5-HT on dorsal-root stimulation-evoked synaptic responses was tested before, during, or after high-frequency conditioning stimulation (CS). In most cells (80%), 5-HT caused a depression of the naı̈ve synaptic response. Even though 5-HT depressed evoked responses, CS in the presence of 5-HT was not only still capable of inducing LTD but also increased its incidence from 54% in controls to 88% ( P < 0.001). Activation of ligands selective for 5-HT1A/1B and 5-HT1B, but not 5-HT2A/2C or 5-HT3receptors, best reproduced these actions. 5-HT also potently depressed postconditioning synaptic responses regardless of whether the induced plasticity was LTP or LTD. Our results demonstrate that in addition to depressing the amplitude of evoked sensory input, 5-HT can also control the direction of its long-term modifiability, favoring the expression of LTD. These findings demonstrate cellular mechanisms that may contribute to the descending serotonergic control of nociception.


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