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D D Samulack

Publications and source records attributed to D D Samulack.

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Membrane properties and synaptic responses of interneurons located near the stratum lacunosum-moleculare/radiatum border of area CA1 in whole-cell recordings from rat hippocampal slices.

1. The membrane properties and synaptic inputs of interneurons, located at the stratum (s.) lacunosum-moleculare and radiatum border (L-M) of the CA1 region, were examined with the use of current-clamp whole-cell recordings in rat hippocampal slices. 2. Biocytin-labeled L-M interneurons had nonpyramidal somata and aspinous, often beaded, dendrites that arborized in s. lacunosum-moleculare and radiatum, sometimes as far as s. moleculare of the dentate gyrus. Their axon coursed and branched in s. lacunosum-moleculare and radiatum. Axon collaterals were also observed traversing the hippocampal fissure and arborizing in s. moleculare of the dentate gyrus and s. radiatum of the CA3 region. 3. Several membrane properties of interneurons were typically nonpyramidal: they had large input resistances, short-duration action potentials followed by prominent fast afterhyperpolarizations, and responded to hyperpolarizing current pulses with little membrane rectification. L-M interneurons showed significant anodal break responses, and their mean membrane time constant was 33 ms. After-depolarizations elicited by subthreshold depolarizing current pulses were larger in amplitude and decayed more slowly at depolarized than hyperpolarized membrane potentials. 4. The majority of L-M interneurons (35 of 49 cells) were silent at resting membrane potentials, whereas other displayed either spontaneous single action potentials (n = 12) or rhythmic bursts (n = 2). The rhythmic bursts were insensitive to the N-methyl-D-aspartate (NMDA) and non-NMDA excitatory amino acid receptor antagonists, 2-amino-5-phosphonopentanoic acid (AP-5; 50 microM) and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 20 microM), respectively. Both spontaneous single action potentials and burst firing were blocked by membrane hyperpolarization, suggesting that they were intrinsically rather than synaptically generated. 5. L-M interneurons responded with regular sustained firing to depolarizing current pulses at resting membrane potential. However, at more hyperpolarized membrane potentials (near -75 mV), depolarizing current pulses elicited action-potential firing with a delayed onset. This suggests that voltage-sensitive, transient outward currents may be activated in L-M interneurons from hyperpolarized membrane potentials. 6. Electrical stimulation of s. radiatum or lacunosum-moleculare elicited predominantly long-duration excitatory postsynaptic potentials (EPSPs; n = 20 cells), or both EPSPs and inhibitory postsynaptic potentials (IPSPs; n = 17 cells). In most L-M interneurons (35/37), with increasing intensities, up to two action potentials were elicited. Occasionally, larger bursts (3-5 action potentials) were observed (n = 2). 7. The multiphasic components of the synaptic responses became more evident when stimulations were repeated at different membrane potentials.(ABSTRACT TRUNCATED AT 400 WORDS)

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Hyperpolarizing synaptic potentials evoked in CA1 pyramidal cells by glutamate stimulation of interneurons from the oriens/alveus border of rat hippocampal slices. I. Electrophysiological response properties.

To examine the inhibitory postsynaptic potentials (IPSPs) elicited in pyramidal cells by interneurons situated at the stratum oriens/alveus border (O/A), glutamate was applied by micropressure to this area during intracellular recordings from CA1 pyramidal cells. Glutamate stimulation evoked IPSPs (glut-IPSPs) of small amplitude (4 mV), delayed peak latency (100-110 ms), and long duration (300-400 ms). Recurrent activation of interneurons via glutamate stimulation of pyramidal cells by local application in stratum pyramidale (PYR) evoked recurrent IPSPs (PYR glut-IPSPs) with similar amplitude and time course as O/A glut-IPSPs. The mean equilibrium potential of O/A glut-IPSPs (-77 mV) was significantly different from that of the PYR glut-IPSPs (-71 mV), however, neither equilibrium potential was significantly different from that of the electrically evoked early IPSP in the same cells. Glutamate-evoked IPSPs elicited from O/A displayed some response reversal (27% reversal) like those evoked from PYR (41% reversal). The early IPSP evoked by electrical stimulation displayed significantly more response reversal (67% reversal) than glut-IPSPs. Both types of glut-IPSPs (O/A and PYR) were associated with moderate increases in membrane conductance (5.9 and 6.6 nS, respectively), which were significantly less than the conductance change associated with the early IPSP (45.8 nS). In interneurons within PYR, glutamate stimulation in PYR readily elicited a flurry of excitatory postsynaptic potentials, whereas glutamate stimulation in O/A elicited IPSPs. The electrophysiological properties of IPSPs elicited in pyramidal cells by glutamate stimulation of interneurons in O/A were similar to those of recurrent IPSPs evoked from PYR. Given that both of these types of glutamate-evoked IPSPs were mostly mediated via GABAA receptor channels (Samulack DD, Lacaille J-C, 1993, Hippocampus 3:345-358), the small differences observed between equilibrium potentials, response reversals, and conductance changes could be due to a more electronically distant location from the soma of the synapses involved in O/A glut-IPSPs as compared to those of recurrent IPSPs elicited from PYR.

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Hyperpolarizing synaptic potentials evoked in CA1 pyramidal cells by glutamate stimulation of interneurons from the oriens/alveus border of rat hippocampal slices. II. Sensitivity to GABA antagonists.

The receptor type mediating the inhibitory postsynaptic potentials (glut-IPSPs), recorded in CA1 pyramidal cells, as a result of glutamate stimulation of interneurons in stratum oriens near the alveus (O/A) was assessed and compared to the type mediating recurrent IPSPs evoked by recurrent activation of interneurons through glutamate stimulation of pyramidal cells in stratum pyramidale (PYR). In response to repetitive electrical stimulation, the peak amplitude of both the O/A glut-IPSP and the PYR glut-IPSP was attenuated (n = 5) in parallel to the reduction in amplitude of the early and late components of the electrically evoked response (stimulus-evoked disinhibition). This suggested the involvement of GABAergic receptors and attested that the interneurons activated during glut-IPSPs were also involved in the circuitry of the electrically evoked IPSPs. The local application of the selective GABAA antagonist bicuculline (100-200 microM) to the slice resulted in a significant reduction in the amplitude of both the O/A (by 76.5%; n = 9) and PYR (by 86.2%; n = 5) glut-IPSPs, in parallel to a decrease of the electrically evoked early IPSP, but not of the late IPSP. The presence of the GABAB antagonist 2-hydroxy-saclofen (1 mM) was able to significantly reduce the amplitude of the O/A glut-IPSPs (by 27.5%; n = 7) and of the electrically evoked late IPSP, but not the PYR glut-IPSP (n = 3). Although the application of phaclofen (20 mM) to the slice reduced the amplitude of the O/A glut-IPSPs (n = 3), the reduction was not statistically significant. These results suggest that recurrent IPSPs elicited from activation of interneurons by stimulation of pyramidal cells are mediated solely via GABAA receptors. Inhibitory postsynaptic potentials elicited from stimulation of interneurons in O/A were also mediated mostly by GABAA receptors, but in addition, displayed a minor component mediated by GABAB receptors. Therefore, since a large proportion of interneurons in O/A are recurrently excited by pyramidal cells (Lacaille J-C et al., 1987, J Neurosci 7: 1979-1993), and since recurrent IPSPs appeared mediated by GABAA receptors, a subpopulation of interneurons activated from O/A might exist that do not receive recurrent excitation but can inhibit pyramidal cells via GABAB receptors.

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Absence of responses to microstimulation at the hand-face border in baboon primary motor cortex.

Intracortical microstimulation (ICMS) was used to map the primary motor cortex of four adult female baboons, anesthetized with a mixture of halothane and nitrous oxide and supplemented with sodium pentobarbital. The sequence of observed muscle contractions in response to ICMS provided evidence of an orderly mototopic representation of the tongue, face, hand, forearm and upper body. A zone of cortex unresponsive to microstimulation was consistently observed at the border between the face and hand representation of the mototopic map. This zone was observed in all four animals and was consistent over time. Repeated confirmations of the unresponsive nature of these regions were obtained both early and late in the same experiment. No motor-unit responses or muscle contractions were detected by electromyographic (EMG) recording during stimulation of the unresponsive zones. The absence of both visually observed and EMG-recorded contractions and the fact that muscle contractions could be elicited from adjacent regions of cortex with ICMS as low as 1-5 microA provide compelling evidence that the finding reflects a true physiological condition rather than an experimental artifact.

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Topographic organization of baboon primary motor cortex: face, hand, forelimb, and shoulder representation.

(1) The fine details of the motor organization of the forelimb, face, and tongue representation of the baboon (Papio h. anubis) primary motor cortex were studied in four adult animals, using intracortical microstimulation (ICMS). (2) A total of 293 electrode penetrations were made. ICMS was delivered to 10,052 sites, and of these, 6,186 sites were verified to have been located within the grey matter. Motor effects were evoked from 30% of these sites. (3) The baboon motor cortex is confined, in large part, to the cortical tissue lying along the anterior bank of the central sulcus. When the electrode penetrations were confined to the precentral gyrus, few sites were capable of evoking movement when stimulated by currents of 40 microA or less. (4) The details of the motor maps varied among the four animals; nonetheless, a general topographic organization existed, with the tongue musculature being represented most laterally, followed by a medial progression of the face, digits, wrist, forearm, and shoulder. Within the representation of a given body part, the muscles were organized as a mosaic, wherein the same muscle was multiply represented. (5) A zone of unresponsive cortex was observed to lie consistently between the face and forelimb representation in all four animals. Repeated electrode penetrations within the unresponsive zone failed to elicit muscle contractions even with stimulating currents as high as 80 microA. (6) Our results suggest that the baboon motor cortex is topographically organized; however, embedded within this overall pattern lies a fine-grained mosaic incorporating multiple representations of the same muscle.

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Neuroanatomical evidence of reinnervation in primate allografted (transplanted) skin during cyclosporine immunosuppression.

Histological evidence is presented documenting the reinnervation of sensory mechanoreceptors across major histocompatibility barriers in allografted primate (baboon) skin. Meissner and pacinian corpuscles, as well as hair follicles, showed a spectrum of reinnervation by host axons. Our light and electron microscopic evidence to date has suggested that allografted Merkel cells did not survive transplantation. This, and other instances of tissue and receptor destruction resulting from histoincompatibility, indicated marked differences when skin allografts were compared to autografts with respect to the ability of host axons to locate and reinnervate sensory mechanoreceptors.

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Tissue transplants in primates for upper extremity reconstruction: a preliminary report.

Recent advances in clinical transplantation surgery suggest that hand transplantation is no longer an unrealistic expectation. However, two questions must be answered. Can composite tissue transplants survive in a primate species? Does the required neural reinnervation occur under immunosuppression? Four hand transplants and seven neurovascular free flap transplants were done in baboons immunosuppressed with Cyclosporin A and steroids (methylprednisolone). Long-term survival occurred in nine. Electrophysiologic tests of sensory axons revealed reinnervation of transplanted skin as evidenced by well-defined, low threshold receptive fields in the donor tissue. Reinnervation of donor muscle was demonstrated by motor unit recruitment in stepwise fashion after electrical stimulation of the recipient's median and ulnar nerves. Afferent fibers serving the donor's joints and muscle spindles were also observed.

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Experimental models in primates for reconstructive surgery utilizing tissue transplants.

Two experimental models for tissue transplantation between unrelated individuals of a primate species have been designed to study survival and reinnervation. The first is a neurovascular free flap consisting of the entire soft tissue coverage of the index finger. The second is an entire hand transplant through the distal forearm. Ongoing studies show that cyclosporin A at high doses, in combination with a tapering regimen of steroids to a low maintenance level, permits prolonged survival of both transplant models. Careful biochemical, hematological, and cyclosporin A serum trough level monitoring permits use of this drug a very high dosages in primates. Continuing experiments should yield detailed neurophysiological data on the reinnervation of these transplants over the next 6 to 18 months.

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