B. R. Botterman

Carbon nanotube coating improves neuronal recordings

Implanting electrical devices in the nervous system to treat neural diseases is becoming very common. The success of these brain-machine interfaces depends on the electrodes that come into contact with the neural tissue. Here we show that conventional tungsten and stainless steel wire electrodes can be coated with carbon nanotubes using electrochemical techniques under ambient conditions. The carbon nanotube coating enhanced both recording and electrical stimulation of neurons in culture, rats and monkeys by decreasing the electrode impedance and increasing charge transfer. Carbon nanotube-coated electrodes are expected to improve current electrophysiological techniques and to facilitate the development of long-lasting brain-machine interface devices.

Blocking Soluble Tumor Necrosis Factor Signaling with Dominant-Negative Tumor Necrosis Factor Inhibitor Attenuates Loss of Dopaminergic Neurons in Models of Parkinson's Disease

The mechanisms that trigger or contribute to loss of dopaminergic (DA) neurons in Parkinsons disease (PD) remain unclear and controversial. Elevated levels of tumor necrosis factor (TNF) in CSF and postmortem brains of PD patients and animal models of PD implicate this proinflammatory cytokine in the pathophysiology of the disease; but a role for TNF in mediating loss of DA neurons in PD has not been clearly demonstrated. Here, we report that neutralization of soluble TNF (solTNF) in vivo with the engineered dominant-negative TNF compound XENP345 (a PEGylated version of the TNF variant A145R/I97T) reduced by 50% the retrograde nigral degeneration induced by a striatal injection of the oxidative neurotoxin 6-hydroxydopamine (6-OHDA). XENP345 was neuroprotective only when infused into the nigra, not the striatum. XENP345/6-OHDA rats displayed attenuated amphetamine-induced rotational behavior, indicating preservation of striatal dopamine levels. Similar protective effects were observed with chronic in vivo coinfusion of XENP345 with bacterial lipopolysaccharide (LPS) into the substantia nigra, confirming a role for solTNF-dependent neuroinflammation in nigral degeneration. In embryonic rat midbrain neuron/glia cell cultures exposed to LPS, even delayed administration of XENP345 prevented selective degeneration of DA neurons despite sustained microglia activation and secretion of solTNF. XENP345 also attenuated 6-OHDA-induced DA neuron toxicity in vitro. Collectively, our data demonstrate a role for TNF in vitro and in vivo in two models of PD, and raise the possibility that delaying the progressive degeneration of the nigrostriatal pathway in humans is therapeutically feasible with agents capable of blocking solTNF in early stages of PD.

Localization of monosynaptic Ia excitatory post‐synaptic potentials in the motor nucleus of the cat biceps femoris muscle.

Evidence is presented for the existence of a localization of monosynaptic Ia excitatory post‐synaptic potentials (e.p.s.p.s) in the motor nucleus of a cat hind limb muscle. Intracellular recordings from biceps femoris motoneurones were made in anaesthetized low spinal cats of the effects of stimuli to the nerve branches supplying the anterior, middle, and posterior portions of the biceps femoris muscle. Recordings were also made during stimulation of nerves to semimembranosus and semitendinosus in order to provide a means of categorizing middle biceps cells as ‘extensors’ (middle biceps‐extensor; i.e. like anterior biceps cells) or as ‘flexors’ (middle biceps‐flexor; like posterior biceps). Homonymous nerve‐branch (i.e. from anterior, middle or posterior biceps) monosynaptic Ia e.p.s.p.s were compared within unifunctional (flexor or extensor) groups of motoneurones. In three of four comparisons (anterior biceps nerve branch onto anterior and middle biceps‐extensor cells, middle biceps onto middle biceps‐flexor and posterior biceps, posterior biceps onto middle biceps‐flexor and posterior biceps) the anterior, middle and posterior biceps nerve branches contributed larger e.p.s.p.s to their ‘own’ motoneurones than to motoneurones supplying other ‘compartments’ of the muscle. In the fourth case, middle bicepss input appeared to have similar effects onto anterior biceps and middle biceps‐extensor cells. A normalization was performed to eliminate the possibility that the differences in e.p.s.p. sizes were due to differences in cell type within the four cell groupings (i.e. differences in the number of cells supplying FF, F(int.), FR and S muscle units). This normalization confirmed that the localization in the first three comparisons was not a consequence of differences in motoneurone type and, in addition, suggested that middle biceps may indeed have greater effects on middle biceps‐extensor than anterior biceps cells. In addition to the asymmetrical effects of anterior and middle biceps nerve branches onto anterior biceps and middle biceps‐extensor motoneurones, it was shown that while semitendinosus and posterior biceps contributed larger e.p.s.p.s to middle biceps‐flexor than to middle biceps‐extensor cells, the anterior biceps nerve branch and semimembranosus nerve contributed equally to the two middle biceps groups. Analysis of cell location in the spinal cord and rostro‐caudal differences in group I volley sizes gave evidence of a topographic organization of the biceps femoris motor nucleus which could contribute to the observed localization. However, localization was also evident when comparing e.p.s.p. amplitudes in pairs of neighbouring cells of different category, indicating a role for neuronal recognition factors.

Distribution of monosynaptic Ia excitatory post‐synaptic potentials in the motor nucleus of the cat semitendinosus muscle.

Evidence is presented for a lack of localization of monosynaptic Ia excitatory post‐synaptic potentials (e.p.s.p.s) in the motor nucleus supplying the atypical cat hind limb muscle semitendinosus, which has anatomically distinct in‐series compartments. Recordings were made from dorsal root filaments containing functionally isolated Ia, spindle group II and Ib axons from the proximal and distal compartments of semitendinosus. Twitch of either of these in‐series compartments resulted in accelerated discharge of Ia and spindle group II fibres in the other compartment. Ib fibres of either compartment showed an in‐series response to twitch of a single compartment which was weaker than twitch of the whole muscle, a finding which was consistent with the diminished force produced by twitch of either compartment alone. In addition, intracellular recordings were made from semitendinosus motoneurones in anaesthetized low‐spinal cats during electrical stimulation of the nerve branches to proximal semitendinosus and distal semitendinosus. Comparison of proximal semitendinosus and distal semitendinosus motoneurones failed to reveal any difference between the two cell groups with respect to the average Ia e.p.s.p. amplitude produced by either the proximal or distal semitendinosus nerve branch. However, e.p.s.p.s due to stimulation of distal semitendinosus were approximately 65% larger, on average, than those due to stimulation of proximal semitendinosus in either motoneurone group. Analysis of cell location along the rostro‐caudal axis of the spinal cord indicated that the proximal and distal semitendinosus cell groups are largely co‐extensive. Recordings of volleys in the proximal and distal semitendinosus nerve branches in response to stimulation of the L6, L7 and S1 dorsal roots showed that group I afferents from the proximal semitendinosus compartment tend to have a more rostral entry point to the spinal cord than do distal semitendinosus afferents. E.p.s.p. amplitude in either cell group due to stimulation of either nerve branch showed little dependence on cell location in the spinal cord. The results are discussed with respect to the relation between muscle function and the distribution of monosynaptic Ia connexions.

Autologous transplants of Adipose-Derived Adult Stromal (ADAS) cells afford dopaminergic neuroprotection in a model of Parkinson's disease

Adult adipose contains stromal progenitor cells with neurogenic potential. However, the stability of neuronal phenotypes adopted by Adipose-Derived Adult Stromal (ADAS) cells and whether terminal neuronal differentiation is required for their consideration as alternatives in cell replacement strategies to treat neurological disorders is largely unknown. We investigated whether in vitro neural induction of ADAS cells determined their ability to neuroprotect or restore function in a lesioned dopaminergic pathway. In vitro-expanded naïve or differentiated ADAS cells were autologously transplanted into substantia nigra 1 week after an intrastriatal 6-hydroxydopamine injection. Neurochemical and behavioral measures demonstrated neuroprotective effects of both ADAS grafts against 6-hydroxydopamine-induced dopaminergic neuron death, suggesting that pre-transplantation differentiation of the cells does not determine their ability to survive or neuroprotect in vivo. Therefore, we investigated whether equivalent protection by naïve and neurally-induced ADAS grafts resulted from robust in situ differentiation of both graft types into dopaminergic fates. Immunohistological analyses revealed that ADAS cells did not adopt dopaminergic cell fates in situ, consistent with the limited ability of these cells to undergo terminal differentiation into electrically active neurons in vitro. Moreover, re-exposure of neurally-differentiated ADAS cells to serum-containing medium in vitro confirmed ADAS cell phenotypic instability (plasticity). Lastly, given that gene expression analyses of in vitro-expanded ADAS cells revealed that both naïve and differentiated ADAS cells express potent dopaminergic survival factors, ADAS transplants may have exerted neuroprotective effects by production of trophic factors at the lesion site. ADAS cells may be ideal for ex vivo gene transfer therapies in Parkinsons disease treatment.

Motor unit - muscle spindle interactions in active muscles of decerebrate cats

Single muscle spindle afferent and motor unit EMG spike trains have been recorded simultaneously during periods of spontaneous motor activity in triceps surae muscles of decerebrate cats. The approximate time course and magnitude of the motor unit contractions were extracted from the whole muscle force record by spike-triggered averaging, and the functional interactions between motor unit contractions and spindle discharge were assessed by cross-correlating their respective spike trains. We have found that both spindle group Ia and II afferents are responsive to the contractions of single motor units in the presence of spontaneous motor activity, being strongly coupled to the activity of some motor units and indifferent to the contractions of others. Moreover, the cross-correlation analysis revealed modulation of a single motor units discharge pattern by the input of a single Ia afferent.

Effects of lateral reticular nucleus lesions on the exercise pressor reflex in cats

Electrical stimulation of ventral roots gives rise to a reflex cardiovascular reponse similar to that observed during static exercise. Although the afferent limb of the reflex is known to be comprised of small diameter afferent fibers from the contracting muscle, little is known of the central nervous system pathway(s) involved. The lateral reticular nucleus of the brainstem is known to be an important site of integration for numerous types of visceral and somatic afferent information, many of which give rise to cardiovascular responses. However, the linkage between the small diameter muscle afferents responsible for the exercise pressor reflex and the lateral reticular nucleus has not been established. In anesthetized cats (n = 7), stimulation of L7 and Si ventral roots increased mean arterial pressure (18.6 ± 2.4 mm Hg) and heart rate (7.4 ± 1.7 beats/min). Following bilateral lesions of the lateral reticular nucleus, the increases in mean arterial pressure and in heart rate were essentially abolished (P < 0.005) (mean arterial pressure increased 1.9 ± 0.8 mm Hg and heart rate increased 0.7 ± 0.5 beats/min). Unilateral lateral reticular nucleus lesions and control lesions in pressor sites outside the lateral reticular nucleus (n = 5) did not affect the exercise pressor reflex. The lateral reticular nucleus lesions also produced decreases (P < 0.01) both in resting mean arterial pressure (—27 ± 5.5 mm Hg) and heart rate (—31.0 ± 8 beats/min). These data suggest that the lateral reticular nucleus is important in the central pathway of the exercise pressor reflex and mediates a tonic pressor influence at rest. (Circ Res 51: 4OO-403, 1982)

Early interfaced neural activity from chronic amputated nerves

Direct interfacing of transected peripheral nerves with advanced robotic prosthetic devices has been proposed as a strategy for achieving natural motor control and sensory perception of such bionic substitutes, thus fully functionally replacing missing limbs in amputees. Multi-electrode arrays placed in the brain and peripheral nerves have been used successfully to convey neural control of prosthetic devices to the user. However, reactive gliosis, micro hemorrhages, axonopathy and excessive inflammation currently limit their long-term use. Here we demonstrate that enticement of peripheral nerve regeneration through a non-obstructive multi-electrode array, after either acute or chronic nerve amputation, offers a viable alternative to obtain early neural recordings and to enhance long-term interfacing of nerve activity. Non-restrictive electrode arrays placed in the path of regenerating nerve fibers allowed the recording of action potentials as early as 8 days post-implantation with high signal-to-noise ratio, as long as 3 months in some animals, and with minimal inflammation at the nerve tissue-metal electrode interface. Our findings suggest that regenerative multi-electrode arrays of open design allow early and stable interfacing of neural activity from amputated peripheral nerves and might contribute towards conveying full neural control and sensory feedback to users of robotic prosthetic devices.

Metabolic and contractile changes in fast and slow muscles of the cat after glucocorticoid-induced atrophy

Abstract The susceptibility of fast- and slow-twitch hind limb muscles to glucocorticoid-induced atrophy was investigated in adult male cats treated for 10 to 14 days with triamcinolone (4 mg/kg/day), using several histochemical, biochemical, and functional indices. After treatment, muscle weight loss in the fast-twitch muscles (medial gastrocnemius and vastus medialis) occurred to a greater extent than in the slow-twitch muscles (soleus and vastus intermedius), with the latter muscles decreasing in weight proportional to the body weight. Fast-twitch glycolytic (FG) fibers responded with similar degrees of atrophy in the muscles examined; however, slow-twitch oxidative (SO) and fast-twitch oxidative glycolytic (FOG) fibers atrophied more in the fast-twitch compared to the slow-twitch muscles. Phosphofructokinase and NADP+-linked isocitrate dehydrogenase specific activities decreased similarly in the fast-twitch muscles, while no change occurred in the slow-twitch muscles. Functionally, the soleus and medial gastrocnemius remained unchanged in abilities to generate tension tetanically, when this was expressed per unit muscle mass or per unit contractile protein. As a result of the treatment, however, the medial gastrocnemius fatigued faster in response to repetitive stimulation in the glucocorticoid-treated animals. The results suggest that the response of muscle to glucocorticoid-induced atrophy is not regulated by the primary metabolic pathways used for energy production. The differences in response of the SO and FOG fiber types in fast- versus slow-twitch muscles suggest basic differences in metabolic and activity profiles of the same fiber types in different muscles, which may influence susceptibility to atrophy.

Activation of caudal brainstem cell groups during the exercise pressor reflex in the cat as elucidated by 2-[14C]deoxyglucose

Cell groups of the caudal brainstem were labeled with 2-[14C]deoxyglucose during the pressor response evoked by contraction of hindlimb muscles (exercise pressor reflex). The nuclear groups which were labeled in excess of control levels included: the lateral reticular nucleus, the inferior olive (medial accessory olive), and the lateral tegmental field (adjacent to the lateral reticular nucleus).