Functional connectivity in the rodent trigeminal pathway grown in vitro.
In explant cocultures of the rat trigeminal pathway, embryonic trigeminal ganglion cells grow their axons into peripheral cutaneous and central nervous system targets (R.S. Erzurumlu, S. Jhaveri, Target influences on the morphology of trigeminal axons, Exp. Neurol, 135 (1995) 1-16; R.S. Erzurumlu, S. Jhaveri, H. Takahashi, R.D.G. McKay, Target-derived influences on axon growth modes in explant cocultures of trigeminal neurons, Proc. Natl. Acad. Sci. USA 90 (1993) 7235-7239). In heterochronic cocultures, composed of embryonic trigeminal ganglion, embryonic whisker pad and postnatal brainstem slice, trigeminal axons develop arbors and terminal boutons in the brainstem trigeminal nuclei. To determine whether these terminal arbors establish functional connections with the brainstem neurons, we examined the electrophysiological properties of brainstem neurons and their responsiveness to trigeminal ganglion stimulation. Intracellular recordings were done in vitro on cells of the trigeminal subnucleus interpolaris (SPI) in trigeminal pathway cocultures (E15 whisker pad, E15 trigeminal ganglion, and postnatal day (PND) 0-2 brainstem slice) or in the SPI of acutely prepared brainstem slices. Electrophysiological properties of SPI cells in both preparations were virtually identical. The voltage responses of SPI neurons to intracellular current injection were highly linear suggesting they lacked a number of voltage-dependent conductances. Depolarizing current injection produced trains of action potentials with a frequency that varied with stimulus intensity. In explant cocultures, electrical activation of the trigeminal ganglion evoked EPSPs, and EPSPs coupled with IPSPs in SPI cells. Bicuculline blockade of IPSP activity resulted in long lasting EPSPs whose duration increased with membrane depolarization. These results show that brainstem trigeminal neurons can retain their functional properties in culture and establish functional connections with primary sensory afferents.
Erzurumlu
R S
RS
Department of Anatomy, LSU Medical Center, New Orleans 70112, USA. RERZUR@LSUMC.EDU
Lo
F S
FS
Günhan-Agar
E
E
Guido
W
W
D000818:Animals; D001369:Axons / Q000502:physiology / Q000648:ultrastructure; D002450:Cell Communication / Q000502:physiology*; D004594:Electrophysiology; D005260:Female; D008564:Membrane Potentials / Q000502:physiology; D009434:Neural Pathways / Q000166:cytology / Q000254:growth & development; D009924:Organ Culture Techniques; D011247:Pregnancy; D051381:Rats; D017207:Rats, Sprague-Dawley; D013570:Synaptic Membranes / Q000378:metabolism / Q000502:physiology; D016707:Tissue Fixation; D012668:Trigeminal Ganglion / Q000166:cytology / Q000254:growth & development; D014276:Trigeminal Nerve / Q000166:cytology / Q000254:growth & development*; D014278:Trigeminal Nuclei / Q000166:cytology / Q000254:growth & development