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Hindfoot, midfoot, and forefoot motion during the stance phase of walking provide insights into the forward progression of the body over the feet via the rocker mechanisms. These segmental motions are affected by walking speed. Increases in walking speed are accomplished by increasing step length and cadence. It is unknown if taking short, medium, and long steps at the same speed would increase hindfoot, midfoot, and forefoot motion similarly to walking speed. We examined effects of different step lengths at the same preferred walking speed on peak forefoot, midfoot, and hindfoot motions related to the foot rockers. Twelve young healthy adults completed five walking trials under three step length conditions in a random order as feet and lower extremity motion were measured via marker positions for the combined Oxford foot and conventional gait models. Peak hindfoot, midfoot, and forefoot joint angles indicating heel, ankle, and forefoot rockers were identified. When walking at the same preferred speed with increase in step length, there were increases in peak hindfoot-tibia plantarflexion angle (p < 0.001; η p 2  = 0.76) in early stance associated with the heel rocker and peak hindfoot-tibia dorsiflexion angle (p = 0.016; η p 2  = 0.39) in midstance associated with ankle rocker. In late stance, the peak hindfoot-tibia plantarflexion angle, forefoot-hindfoot angle, and forefoot-hallux dorsiflexion angle indicating forefoot rocker motion also increased with step length (p < 0.01). When foot kinematics are compared across different individuals or the same individual across different sessions, researchers and clinicians should consider the influence of step length as a contributor to differences in foot kinematics observed., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Published by Elsevier Ltd.)

Background: Hallux valgus (HV) contributes to deficits in static balance and increased fall risk in older adults. Very limited research has examined dynamic balance deficits in walking in this population. These individuals generally walk slowly, as balance challenge is lesser at slow speeds., Research Question: How does the dynamic balance of older adults with HV differ from healthy controls at controlled slow and fast walking speeds?, Methods: Nineteen older adults with HV and 13 healthy controls completed 5 continuous walking trials at 1.0 and 1.3 m·s -1 as whole body marker position and ground reaction force data were captured. Dynamic balance was evaluated using whole body center of mass (COM) and center of pressure (COP) inclination angles (IA) and duration of double support., Results: There were no differences in measures of dynamic balance between older adults with and without HV at slow and fast speeds. At the faster speed, the peak sagittal plane COM-COP IA increased and the double support duration decreased, while the peak frontal plane COM-COP IA were not affected., Significance: Older adults with HV do not exhibit deficits in dynamic balance during continuous walking at comfortable speeds when compared to healthy older adults., Competing Interests: Declaration of Competing Interest The authors have no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Characterization of proteins using NMR methods begins with assignment of resonances to specific residues. This is usually accomplished using sequential connectivities between nuclear pairs in proteins uniformly labeled with NMR active isotopes. This becomes impractical for larger proteins, and especially for proteins that are best expressed in mammalian cells, including glycoproteins. Here an alternate protocol for the assignment of NMR resonances of sparsely labeled proteins, namely, the ones labeled with a single amino acid type, or a limited subset of types, isotopically enriched with 15 N or 13 C, is described. The protocol is based on comparison of data collected using extensions of simple two-dimensional NMR experiments (correlated chemical shifts, nuclear Overhauser effects, residual dipolar couplings) to predictions from molecular dynamics trajectories that begin with known protein structures. Optimal pairing of predicted and experimental values is facilitated by a software package that employs a genetic algorithm, ASSIGN&#95;SLP&#95;MD. The approach is applied to the 36-kDa luminal domain of the sialyltransferase, rST6Gal1, in which all phenylalanines are labeled with 15 N, and the results are validated by elimination of resonances via single-point mutations of selected phenylalanines to tyrosines. Assignment allows the use of previously published paramagnetic relaxation enhancements to evaluate placement of a substrate analog in the active site of this protein. The protocol will open the way to structural characterization of the many glycosylated and other proteins that are best expressed in mammalian cells., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Reduced stability while standing typically decreases the soleus muscle Hoffmann (H-) reflex amplitude, purportedly to prevent the Ia afferent signal from excessively activating spinal motor neurons during the unstable stance. H-reflex measures, however, by excluding the spindle do not reflect the actual effect of the Ia pathway (i.e. the combined effects of spindle sensitivity and Ia presynaptic inhibition) on motor neuron activation, as tendon tap reflex measures can. But the effect of stance stability on soleus muscle tendon tap reflex amplitude is largely unknown. This study examined 30 young adults (mean(s), 21(2) years) as they stood in a wide stable stance position and an unstable tandem stance with a reduced base of support. Standing body sway, the amplitude of the soleus muscle tendon tap reflex, background EMG and tap force were measured in both stances. A repeated measured design t-test was calculated for each variable. Most subjects (69%) decreased tendon tap reflex amplitude when in the tandem stance position (mean decrease 11.6%), compared to the wide stance (wide stance 0.248(0.124) mV, tandem stance 0.219(0.119) mV, p < 0.05, Cohen's d = 0.24 small) with no significant differences in background soleus and tibialis anterior EMG, and tap force across the stances. There was no relationship between the modulation of the tendon tap reflex amplitude across the stances and standing body sway in the tandem stance. Results support the idea that for most subjects examined, during a less stable stance the Ia excitation of motor neurons is decreased, likely by presynaptic inhibition, thereby avoiding potential instability in the reflex loop or saturating the reflex pathway and possibly interfering with descending control of the involved spinal motor neurons., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Gilmore, SL, Brilla, LR, Suprak, DN, Chalmers, GR, and Dahlquist, DT. Effect of a high-intensity isometric potentiating warm-up on bat velocity. J Strength Cond Res 33(1): 152-158, 2019-This study examined the acute effect of a high-intensity isometric potentiating warm-up on subsequent maximal horizontal bat velocity in experienced female softball players (n = 28). The isometric potentiating warm-up consisted of 3 sets of 5-second maximal voluntary contractions held in the early swing phase position, pulling against an immovable device. As one of the simplest methods to use the postactivation potentiation (PAP) stimulus, the warm-up was designed to acutely enhance muscle performance by inducing PAP. Because optimal recovery duration after a potentiating warm-up can be highly variable, swing trials were conducted at predetermined rest intervals (1, 2, 4, 6, 8, 10, and 12 minutes) to identify the recovery time that may have allowed for maximal possible benefits. Bat velocity was measured immediately before bat-ball impact using MaxTRAQ motion analysis software. The results showed that maximal horizontal bat velocity was significantly enhanced 6 minutes after the isometric warm-up protocol (+1.27 m·s, +2.84 mph, +4.93%; p < 0.05). In experienced female softball athletes, a specific isometric warm-up may acutely enhance maximal horizontal bat velocity.

Background: Creatine supplementation is recommended as an ergogenic aid to improve repeated sprint cycling performance. Furthermore, creatine uptake is increased in the presence of electrolytes. Prior research examining the effect of a creatine-electrolyte (CE) supplement on repeated sprint cycling performance, however, did not show post-supplementation improvement. The purpose of this double blind randomized control study was to investigate the effect of a six-week CE supplementation intervention on overall and repeated peak and mean power output during repeated cycling sprints with recovery periods of 2 min between sprints., Methods: Peak and mean power generated by 23 male recreational cyclists (CE group: n  = 12; 24.0 ± 4.2 years; placebo (P) group: n  = 11; 23.3 ± 3.1 years) were measured on a Velotron ergometer as they completed five 15-s cycling sprints, with 2 min of recovery between sprints, pre- and post-supplementation. Mixed-model ANOVAs were used for statistical analyses., Results: A supplement-time interaction showed a 4% increase in overall peak power (pre: 734 ± 75 W; post: 765 ± 71 W; p  = 0.040; η p 2  = 0.187) and a 5% increase in overall mean power (pre: 586 ± 72 W; post: 615 ± 74 W; p  = 0.019; η p 2  = 0.234) from pre- to post-supplementation for the CE group. For the P group, no differences were observed in overall peak (pre: 768 ± 95 W; post: 772 ± 108 W; p  = 0.735) and overall mean power (pre: 638 ± 77 W; post: 643 ± 92 W; p  = 0.435) from pre- to post-testing. For repeated sprint analysis, peak (pre: 737 ± 88 W; post: 767 ± 92 W; p  = 0.002; η p 2  = 0.380) and mean (pre: 650 ± 92 W; post: 694 ± 87 W; p  < 0.001; η p 2  = 0.578) power output were significantly increased only in the first sprint effort in CE group from pre- to post-supplementation testing. For the P group, no differences were observed for repeated sprint performance., Conclusion: A CE supplement improves overall and repeated short duration sprint cycling performance when sprints are interspersed with adequate recovery periods., Competing Interests: The study design and procedures were approval by the Western Washington University Institutional Review Board. All subjects signed a written informed form prior to participating in the study.The authors declare that they have no competing interests.Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

A strategy for acquiring structural information from sparsely isotopically labeled large proteins is illustrated with an application to the E. coli heat-shock protein, HtpG (high temperature protein G), a 145 kDa dimer. It uses 13 C-alanine methyl labeling in a perdeuterated background to take advantage of the sensitivity and resolution of Methyl-TROSY spectra, as well as the backbone-centered structural information from 1 H- 13 C residual dipolar couplings (RDCs) of alanine methyl groups. In all, 40 of the 47 expected crosspeaks were resolved and 36 gave RDC data. Assignments of crosspeaks were partially achieved by transferring assignments from those made on individual domains using triple resonance methods. However, these were incomplete and in many cases the transfer was ambiguous. A genetic algorithm search for consistency between predictions based on domain structures and measurements for chemical shifts and RDCs allowed 60% of the 40 resolved crosspeaks to be assigned with confidence. Chemical shift changes of these crosspeaks on adding an ATP analog to the apo-protein are shown to be consistent with structural changes expected on comparing previous crystal structures for apo- and complex- structures. RDCs collected on the assigned alanine methyl peaks are used to generate a new solution model for the apo-protein structure.

Sparse isotopic labeling of proteins for NMR studies using single types of amino acid ( 15 N or 13 C enriched) has several advantages. Resolution is enhanced by reducing numbers of resonances for large proteins, and isotopic labeling becomes economically feasible for glycoproteins that must be expressed in mammalian cells. However, without access to the traditional triple resonance strategies that require uniform isotopic labeling, NMR assignment of crosspeaks in heteronuclear single quantum coherence (HSQC) spectra is challenging. We present an alternative strategy which combines readily accessible NMR data with known protein domain structures. Based on the structures, chemical shifts are predicted, NOE cross-peak lists are generated, and residual dipolar couplings (RDCs) are calculated for each labeled site. Simulated data are then compared to measured values for a trial set of assignments and scored. A genetic algorithm uses the scores to search for an optimal pairing of HSQC crosspeaks with labeled sites. While none of the individual data types can give a definitive assignment for a particular site, their combination can in most cases. Four test proteins previously assigned using triple resonance methods and a sparsely labeled glycosylated protein, Robo1, previously assigned by manual analysis, are used to validate the method and develop a criterion for identifying sites assigned with high confidence.

Objective: Investigate the effects of shoulder elevation on repositioning errors in upright and supine body orientations, and examine these effects on anterior and posterior deltoid muscle activation. We hypothesized decreased errors, and altered anterior and posterior deltoid activation with increasing elevation, in both orientations., Design: Crossover trial., Setting: University laboratory., Participants: Thirty-five college-aged participants., Intervention: Subjects attempted to replicate target positions of various elevation angles in upright and supine body orientations. Also, anterior and posterior deltoid activation was recorded in each shoulder position and body orientation., Main Outcome Measures: Vector and variable repositioning errors, anterior and posterior deltoid percentage of maximal contraction., Results: Vector error was greater in supine compared to upright at 90° and 110°, but not at 70°. Variable error was larger in supine than upright, but was unaffected by elevation. Anterior deltoid activation increased with elevation in the upright posture only. Posterior deltoid activation increased with elevation across postures., Conclusions: Muscle activation, external torque, and cutaneous sensations may combine to provide afferent feedback, and be used with centrally-generated signals to interpret the state of the limb during movement. Clinicians may prescribe open kinetic chain exercises in the upright posture with the shoulder elevated approximately 90-100°., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Context: Joint position sense (JPS) is a key neuromuscular factor for developing and maintaining control of muscles around a joint. It is important when performing specialized tasks, especially at the shoulder. No researchers have studied how Kinesio Tape (KT) application affects JPS., Objective: To investigate the effects of KT application and no tape on shoulder JPS at increasing shoulder elevations in athletes., Design: Cross-sectional study., Setting: University laboratory., Patients or Other Participants: A total of 27 healthy athletes who did not participate in overhead sports (age = 20.44 ± 1.05 years, height = 175.02 ± 11.67 cm, mass = 70.74 ± 9.65 kg) with no previous pathologic shoulder conditions volunteered for the study. All participants were from 1 university., Intervention(s): Shoulder JPS was assessed at increasing elevations with and without KT application. Participants attempted to actively replicate 3 target positions with and without the KT and without visual guidance., Main Outcome Measure(s): We examined absolute and variable repositioning errors at increasing shoulder-elevation levels with and without KT application., Results: Data revealed an interaction between tape and position for absolute error (F2,52 = 4.07, P = .02); simple effects revealed an increase in error, with KT demonstrating a 2.65° increase in error at 90° of elevation compared with no tape (t26 = 2.65, P = .01). The effect size was medium (ω(2) = .135). Variable error showed no interaction of tape and position (F2,52 = .709, P = .50). Further analysis of simple effects was not needed. However, we still calculated the effect size and observed small effect sizes for tape (ω(2) = .002), position (ω(2) = .072), and tape by position (ω(2) = .027)., Conclusions: At 90° of elevation, shoulder JPS was impaired by the application of KT.

Exercise science and human anatomy and physiology textbooks commonly report that type IIB muscle fibers have the largest cross-sectional area of the three fiber types. These descriptions of muscle fiber sizes do not match with the research literature examining muscle fibers in young adult nontrained humans. For men, most commonly type IIA fibers were significantly larger than other fiber types (six out of 10 cases across six different muscles). For women, either type I, or both I and IIA muscle fibers were usually significantly the largest (five out of six cases across four different muscles). In none of these reports were type IIB fibers significantly larger than both other fiber types. In 27 studies that did not include statistical comparisons of mean fiber sizes across fiber types, in no cases were type IIB or fast glycolytic fibers larger than both type I and IIA, or slow oxidative and fast oxidative glycolytic fibers. The likely reason for mistakes in textbook descriptions of human muscle fiber sizes is that animal data were presented without being labeled as such, and without any warning that there are interspecies differences in muscle fiber properties. Correct knowledge of muscle fiber sizes may facilitate interpreting training and aging adaptations.

Mirrors are often used in an instructional environment where precise movements must be learned (e.g., martial arts, Pilates, dance). The potential for mirrors in the learning environment of a Pilates class, to affect the subsequent performance of a Pilates star movement when mirrors are not present, was examined. Twenty subjects learned the Pilates star movement over seven weeks, either with (n=11) or without (n=9), mirrors present in the Pilates studio. Performance of the star without mirrors present was assessed quantitatively before and after the training, by video analysis of the degree of lateral straightness of the subject's body at the start, middle, and end of the star movement. Performance of the star movement without a mirror present improved similarly for both the group that learned with, and the group that learned without, mirrors present (p<0.05). These results indicate that the inclusion of mirrors in a learning environment, to provide immediate visual feedback during learning, does not necessarily enhance the subsequent performance of a skill when mirrors are not present.

Literature examining the recruitment order of motor units during lengthening (eccentric) contractions was reviewed to determine if fast-twitch motor units can be active while lower threshold slow-twitch motor units are not active. Studies utilizing surface electromyogram (EMG) amplitude, single motor unit activity, spike amplitude-frequency analyses, EMG power spectrum, mechanomyographic, and phosphocreatine-to-creatine ratio (PCr/Cr) techniques were reviewed. Only single motor unit and PCr/Cr data were found to be suitable to address the goals of this review. Nine of ten single motor unit studies, examining joint movement velocities up to 225 degrees/s and forces up to 53% of a maximum voluntary contraction, found that the size principle of motor unit recruitment applied during lengthening contractions. Deviation from the size principle was demonstrated by one study examining movements within a small range of low velocities and modest forces, although other studies examining similar low forces and lengthening velocities reported size-ordered recruitment. The PCr/Cr data demonstrated the activation of all fibre types in lengthening maximal contractions. Most evidence indicates that for lengthening contractions of a wide range of efforts and speeds, fast-twitch muscle fibres cannot be selectively recruited without activity of the slow-twitch fibres of the same muscle.

Health professionals and members of the public are often interested in locating exercise information on the World Wide Web (the Web). There is a large amount of information available on the Web; however, the challenge for all people is to identify the high quality information that can be depended upon. Much of the quality exercise information on the Web is hidden within sites of reputable organizations concerned with exercise and health. This article examines several categories of commonly needed exercise related information. For each category, a few high quality sources of exercise information are listed. Combined, these Web sites provide an excellent and extensive body of knowledge for a person who is not experienced with exercise and wants to get started learning, or for a person with a moderate level of experience and knowledge who wants to learn more.

Recurrent inhibition in the spinal cord has been suggested to serve as a variable gain regulator to allow for optimal muscle force control, to influence alpha-motoneuron firing rate, and to contribute to task related motor synergies between muscles at the same or different joints. The purpose of this study was to examine the resting recurrent inhibition levels in the soleus motoneuron pool of 20 elderly and 21 young adult subjects. To assess recurrent inhibition, a conditioning electrical stimulus was used to activate group Ia afferent fibers and elicit a reflex response in some of the a-motoneurons innervating the soleus muscle; producing both activation of Renshaw interneurons excited by those involved soleus a-motoneurons via a recurrent branch of the a-motoneuron axon, and an H-reflex response in the soleus muscle. A H' test reflex elicited by a successive supramaximal stimulus to the same nerve 10 ms after the conditioning stimulus evaluated the resulting inhibitory effect. There was no difference in the H' test reflex amplitude when comparing the young and elderly adult subjects. This result was found following two different methods employed to control for a possible effect on the H' test reflex amplitude of a smaller maximum H-reflex amplitude in the elderly subjects. These results indicate that the level of recurrent inhibition in the motoneuron pool of the resting soleus muscles of the young and elderly adults examined was not significantly different.

Soleus Hoffman-reflex (H-reflex) gain was compared at the same background level of electromyographic activity across lying, natural standing, and tandem stance postures, in 12 young and 16 elderly adults. When compared to a lying posture, young adults significantly depressed soleus H-reflex gain when in a natural standing (19% decrease) and a tandem stance position (30% decrease; p <.0125 for both positions). For elderly adults, there was no significant decrease in H-reflex gain while standing naturally, but there was a significant 28% decrease when performing tandem stance (p <.0125). The data indicate that, although the mild motor control challenge of natural standing does not induce a decrease in soleus H-reflex gain in the elderly adults, as it does in young adults, in the more difficult task of tandem stance, soleus H-reflex gain is significantly decreased in both young and elderly adults.

Transcranial magnetic stimulation (TMS) of the motor cortex excites limb muscles of the contralateral side of the body. Reports of poorly defined, or a complete lack of systematic excitatory responses of soleus motoneurons compared with those of tibialis anterior (TA) motoneurons has led to the proposal that while all ankle flexor motoneurons receive strong corticomotoneuronal connections, very few soleus motoneurons do. In addition, the connections to these few motoneurons are weak. The nature of corticomotoneuronal connections onto these two motoneuron pools was re-evaluated in the following experiments. The leg area of the left motor cortex was stimulated with a large double-cone coil using Magstim 200, while surface electromyographic (EMG) and single motor unit (SMU) responses were recorded from soleus and TA muscles of healthy adult subjects. Under resting conditions, the onset (25-30 ms) and duration of concomitantly recorded short latency motor evoked potentials (MEPs) in surface EMG from both muscles were similar. The input-output relationships of the simultaneously recorded soleus and TA EMG responses showed much greater increases in TA MEPs compared with soleus MEPs with identical increases in stimulus intensity. Under resting and nonisometric conditions, a later peak with onset latency of approximately 100 ms was observed in soleus. During isometric conditions or with vibration of the TA tendon, the second soleus peak was abolished indicating reflex origin of this peak. Recordings from 42 soleus and 39 TA motor units showed clear response peaks in the peristimulus time histograms (PSTHs) of every unit. Two statistical tests were done to determine the onset and duration of response peaks in the PSTHs. With chi(2) test, the duration was 6.9 +/- 4.2 ms (mean +/- SD) for soleus and 5.1 +/- 2.1 ms for TA. Using the criterion of discerning a peak by bin counts being three SDs above background, the duration was 10.0 +/- 4.4 ms for soleus and 7.8 +/- 2.6 ms for TA. Results of these experiments do not suggest a lack of systematic corticomotoneuronal connections on soleus motoneurons when compared with those on TA, though some differences in the strengths of corticomotoneuronal connections onto the two pools do exist.

Soleus Hoffmann-reflex (H-reflex) modulation during walking was examined in 7 young and 13 elderly adults. H-reflex size was measured in 16 equal time divisions (phases) of the step cycle. In both the elderly and the young groups, the H reflex was minimal at the time of heel contact, rose to a maximum shortly after midstance, decreased rapidly as toe-off neared, then was minimal during swing. There was a significant interaction between age group and step cycle phase (p < .05). During midstance of walking, the elderly participants had a smaller H-reflex size during two of the 16 time phases of the step cycle (p < .05), despite no significant difference in H-reflex size between the age groups while standing. The smaller H-reflex size during the stance phase of walking may reflect changes in central reflex mechanisms that may impact stretch reflex contribution to ankle extensor neural drive and ankle stiffness in elderly persons during walking.

As one considers changes in motor activity from lower mammals to higher primates, one of the major changes one observes lies in the cortical control of forelimb muscles. There has been a shift from disynaptic control of spinal motoneurons in, for example, the cat, to a greater and greater percentage of monosynaptic control of hand and forelimb motoneurons in the primate. In spite of the species and evolutionary changes in the synaptic connections of the corticospinal tract, it appears that the interneurons identified in the cat are retained in the monkey and human. These interneurons, under the influence of descending pathways, modulate the output of motoneuron pools. Perhaps the control of these interneurons has also changed towards finer control of movement, as has been suggested by recent studies in the monkey. Whether in cat or human, the recruitment pattern for motor units is the same; the change from disynaptic to monosynaptic connections has not changed the recruitment pattern of muscles. Differences in the recruitment patterns of muscles may lie in the finer control of inputs to motoneurons in the primate. This review seeks to integrate the current knowledge of the mechanisms involved in the motor control of the wrist joint and especially in the recruitment patterns of the muscles. These motor control mechanisms include the biomechanics of the wrist joint, recruitment patterns of wrist muscles, interneurons and spinal cord circuits in the cervical regions mediating the output of spinal motoneurons, and the supraspinal control of these muscles.

Short-latency excitatory Ia reflex connections were determined between pairs of human wrist flexor and extensor muscles. Spindle Ia afferents were stimulated by either tendon tap or electrical stimulation. The activity of voluntarily activated single motor units was recorded intramuscularly from pairs of wrist flexor or extensor muscles. Cross-correlation between stimuli and the discharge of the motor units provided a measure of the homonymous or heteronymous excitatory input to a motoneurone. Homonymous motoneurone facilitation was generally stronger than that of the heteronymous motoneurones. The principal wrist flexors, flexor carpi radialis (FCR) and flexor carpi ulnaris (FCU), were tightly connected through a bidirectional short-latency reflex pathway. In contrast, the extensor carpi ulnaris (ECU) and the extensor carpi radialis (ECR) did not have similar connections. ECU motoneurones received no short-latency excitatory Ia input from the ECR. ECR motoneurones did receive excitatory Ia input from ECU Ia afferents; however, its latency was delayed by several milliseconds compared with other heteronymous Ia excitatory effects observed. The wrist and finger extensors were linked through heteronymous Ia excitatory reflexes. The reflex connections observed in humans are largely similar to those observed in the cat, with the exception of heteronymous effects from the ECU to the ECR and from the extensor digitorum communis (EDC) to the ECU, which are present only in humans. The differences in the reflex organization of the wrist flexors versus the extensors probably reflects the importance of grasping.

To assess the cellular and molecular substrates for cholinergic axon growth in the adult central nervous system (CNS), we implanted grafts of control and nerve growth factor (NGF)-producing genetically modified fibroblasts within the striatum of rats. Sprouting cholinergic axonal processes that grew into grafts of NGF-producing fibroblasts were fasciculated and followed the surface of astrocytic processes for long distances within the grafts. The close and long distance anatomical relationship between the sprouted axons and the astrocytes supported previous ultrastructural evidence that astrocytes may serve as a cellular substrate for sprouting cholinergic axons in vivo. The sprouted axon processes were associated with the expression of nerve growth factor-inducible large external (NILE) glycoprotein on their surfaces. NILE expression was not seen in control grafts where there was an absence of cholinergic ingrowth. NILE has been demonstrated to play a role in axon fasciculation in a number of other neural systems. The astrocytic processes in both control and NGF-producing fibroblast grafts expressed neural cell adhesion molecule (NCAM), suggesting that NCAM-mediated adhesion may be responsible for the close relationship between the axons and astrocytes within the grafts. NGF-induced heterotypic interactions between neuronal NILE and astroglial NCAM may also be required for adult cholinergic axonal sprouting.

To determine whether the adaptations in myosin heavy chain (MHC) isoform expression after functional overload (FO) are accompanied by commensurate adaptations in protein isoforms responsible for relaxation [sarco(endo)plasmic reticulum (SR) Ca(2+)-adenosinetriphosphatase (SERCA) and phospholamban (PHL)] in single muscle fibers, the isoforms of MHC and SERCA and the presence or absence of PHL were determined for cat plantaris fibers 3 mo after FO. In control plantaris the relative MHC isoform composition was 23% type I, 21% type IIa, and 56% type IIb. FO resulted in a shift toward slower isoforms (33% type I, 44% type IIa, and 23% type IIb). In the deep region of the plantaris the proportions of type I MHC and hybrid MHC fibers (containing type I and II MHCs) were 40 and 200% greater in FO cats, respectively. FO resulted in a 47% increase in the proportion of fibers containing only the slow SERCA isoform (SERCA2) and a 41% increase in the proportion of fibers containing PHL. The proportions of fibers containing type I MHC, SERCA2, and PHL in control and FO plantaris were linearly correlated. These data show that adaptations in MHC isoform expression are accompanied by commensurate adaptations in sarcoplasmic reticulum protein isoforms in single muscle fibers after FO.

Although central neurons do not naturally recover following injury, damaged adult septal neurons can regenerate when nerve growth factor (NGF) is provided along with a suitable cellular substrate. This study investigates the outgrowth of axotomized septal neurons grafted with primary fibroblasts genetically modified to produce NGF. Confocal microscope images of double staining for neuritic markers (neurofilament or low-affinity NGF receptor) and the astrocytic marker glial fibrillary acidic protein (GFAP) demonstrated that regenerating neurites crossed dense buildups of astrocytic processes at the edges of NGF-producing grafts and were in apposition with astrocytic processes within NGF-producing grafts. Immunoreactivity for acetylcholinesterase and low-(p75) and high-affinity (TrkA) NGF receptors was dense in NGF-producing grafts but absent in control grafts. NGF-grafted rats exhibited significantly increased hippocampal density of p75-immunoreactive fibers and significantly decreased ectopic hippocampal sympathetic ingrowth as compared to control-grafted rats. Rats with unilateral fimbria-fornix lesions and NGF-producing grafts exhibited ameliorated performance on a simple memory task. These findings demonstrate that implantation of NGF-producing grafts to the lesion cavity allows axotomized septal cholinergic neurons to reinnervate the hippocampus, and that rats receiving these grafts show a partial recovery of function.

Chromaffin cells exposed to basic fibroblast growth factor (bFGF) in vitro express characteristics of sympathetic neurons, extend neurites, and become dependent on nerve growth factor (NGF) for survival. We explored whether the growth factor responsiveness of chromaffin cells could be exploited to enhance the transdifferentiation, neurite outgrowth and functional efficacy of chromaffin cells implanted into rats with unilateral 6-hydroxydopamine lesions. Cografts of neonatal chromaffin cells and fibroblasts genetically modified to produce bFGF were placed into the dopamine-depleted striatum of adult rats. Either control-transfected or NGF-producing fibroblasts were then transplanted 1 mm distal to the cograft. Chromaffin cells transdifferentiated under the influence of bFGF, as indicated by the growth of neurites and the expression of neuron-specific proteins. Distal grafts of NGF-producing fibroblasts successfully induced chromaffin neurites to traverse through the host parenchyma to the NGF source. In the absence of NGF fibroblast grafts, neither neurite extension nor good, long-term survival of the chromaffin-derived neurons was observed. Assessments of apomorphine-induced rotational behavior 2- and 4-weeks postgrafting revealed no behavior improvements in any of the groups. These results indicate that localized sources of growth factors are effective in inducing the transdifferentiation of grafted chromaffin cells as well as the extension of chromaffin-derived neurites into the host parenchyma. Such chromaffin cell-derived neurons are, however, functionally ineffective in this rat model of Parkinson's disease. Whether the lack of behavioral improvement reflected the tropic growth of neurites to an inappropriate striatal region or the noradrenergic nature of the chromaffin cell-derived neurons remains to be clarified. Nonetheless, these results caution that promoting transdifferentiation and neurite extension from engrafted chromaffin cells may not be sufficient to achieve desired functional effects of such grafts.

The present study was designed to determine whether the degree and kind of adaptation of a muscle fiber to a functional overload (FO) are determined by properties that are intrinsic to that fiber. The study also addresses the question of the capability of fibers to maintain a normal level of coordination of proteins per fiber as fiber volume changes dramatically. The plantaris muscle of six adult female cats was overloaded for 12 wk by bilateral synergist removal. Plantaris muscle fiber mean size doubled after FO, although some very small fibers that stained dark for adenosinetriphosphatase (ATPase) were observed in some of the FO muscles. There appeared to be no change in total succinate dehydrogenase activity per fiber. A reduction in succinate dehydrogenase activity per unit volume was observed in a substantial number of fibers, reflecting a disproportionate increase in fiber volume relative to mitochondrial volume. In contrast, total alpha-glycerophosphate dehydrogenase activity and actomyosin ATPase activity increased as fiber size increased, whereas there was no change in alpha-glycerophosphate dehydrogenase and ATPase activities per unit volume. Control and FO muscle fibers generally expressed either a fast or slow myosin heavy chain type, but in some cases FO muscle fibers expressed both fast and slow myosin heavy chains. The persistence of variability in fiber sizes and enzyme activities in fibers of overloaded muscles suggests a wide range in the adaptive potential of individual fibers to FO. These data indicate that a severalfold increase in cell size may occur without significant qualitative changes in the coordination of protein regulation associated with metabolic pathways and ATP utilization.

Previous studies have demonstrated that a chronic change in neuronal activation can produce a change in soma oxidative capacity, suggesting that: (i) these 2 variables are directly related in neurons and (ii) ion pumping is an important energy requiring activity of a neuron. Most of these studies, however, have focused on reduced activation levels of sensory systems. In the present study the effect of a chronic increase or decrease in motoneuronal activity on motoneuron oxidative capacity and soma size was studied. In addition, the effect of chronic axotomy was studied as an indicator of whether cytoplasmic volume may also be related to the oxidative capacity of motoneurons. A quantitative histochemical assay for succinate dehydrogenase activity was used as a measure of motoneuron oxidative capacity in experimental models in which chronic electromyography has been used to verify neuronal activity levels. Spinal transection reduced, and spinal isolation virtually eliminated lumbar motoneuron electrical activity. Functional overload of the plantaris by removal of its major synergists was used to chronically increase neural activity of the plantaris motor pool. No change in oxidative capacity or soma size resulted from either a chronic increase or decrease in neuronal activity level. These data indicate that the chronic modulation of ionic transport and neurotransmitter turnover associated with action potentials do not induce compensatory metabolic responses in the metabolic capacity of the soma of lumbar motoneurons. Soma oxidative capacity was reduced in the axotomized motoneurons, suggesting that a combination of axoplasmic transport, intracellular biosynthesis and perhaps neurotransmitter turnover represent the major energy demands on a motoneuron. While soma oxidative capacity may be closely related to neural activity in some neural systems, e.g. visual and auditory, lumbar motoneurons appear to be much less sensitive to modulations in chronic activity levels.

To determine the level of coordination in succinate dehydrogenase (SDH) activity between plantaris motoneurons and muscle fibers, the soleus and gastrocnemius muscles were bilaterally excised in four cats to subject the plantaris to functional overload (FO). Five normal cats served as controls. Twelve weeks after surgery the right plantaris in each cat was injected with horseradish peroxidase to identify plantaris motoneurons. SDH activity then was measured in a population of plantaris motoneurons and muscle fibers in each cat. Control motoneurons and muscle fibers had similar mean SDH activities and a similar relationship between cell size and SDH activity. After FO, muscle fiber size doubled and mean muscle fiber SDH activity halved. Motoneuron mean SDH activity and size were unaffected by FO. Total SDH activity was unchanged in both the motoneurons and muscle fibers after FO. These changes suggest a selective increase in contractile proteins with little or no modulation of mitochondrial proteins in the muscle fibers, because total SDH activity was unchanged in muscle fibers after FO. These data demonstrate that although mean SDH activities were similar in control motoneurons and muscle fibers, mean SDH activities in these two cell types can change independently.

We have developed a quantitative histochemical assay for measurement of succinate dehydrogenase (SDH) activity in single motoneurons. A computer image processing system was used to quantify the histochemical enzyme reaction product and to follow the time course of the reaction. The optimal concentration for each of the ingredients of the incubation medium for the SDH reaction was determined and the importance of using histochemical "blanks" in the determination of enzymatic activity was demonstrated. The enzymatic activity was linear with respect to reaction time and tissue thickness. The procedure described meets the criteria generally considered essential for establishment of a quantitative histochemical assay. The assay was then used to examine the SDH activity of cat and rat motoneurons. It was found that motoneurons with a small soma size had a wide range of SDH activity, whereas those with a large soma size were restricted to low SDH activity.

The effect of tissue fixation on succinate dehydrogenase and cytochrome oxidase activity in single motoneurons of the rat was demonstrated using a computer image processing system. Inhibition of enzyme activity by chemical fixation was variable, with some motoneurons being affected more than others. It was concluded that quantification of enzymatic activity in chemically fixed tissue provides an imprecise estimate of enzyme activities found in fresh-frozen tissues.