Dominant and sensitive control of oxidative flux by the ATP-ADP carrier in human skeletal muscle mitochondria: Effect of lysine acetylation.

The adenine nucleotide translocase (ANT) of the mitochondrial inner membrane exchanges ADP for ATP. Mitochondria were isolated from human vastus lateralis muscle (n = 9). Carboxyatractyloside titration of O2 consumption rate (Jo) at clamped [ADP] of 21 µM gave ANT abundance of 0.97 ±â€¯0.14 nmol ANT/mg and a flux control coefficient of 82% ±â€¯6%. Flux control fell to 1% ±â€¯1% at saturating (2 mM) [ADP]. The KmADP for Jo was 32.4 ±â€¯1.8 µM. In terms of the free (-3) ADP anion this KmADP was 12.0 ±â€¯0.7 µM. A novel luciferase-based assay for ATP production gave KmADP of 13.1 ±â€¯1.9 µM in the absence of ATP competition. The free anion KmADP in this case was 2.0 ±â€¯0.3 µM. Targeted proteomic analyses showed significant acetylation of ANT Lysine23 and that ANT1 was the most abundant isoform. Acetylation of Lysine23 correlated positively with KmADP, r = 0.74, P = 0.022. The findings underscore the central role played by ANT in the control of oxidative phosphorylation, particularly at the energy phosphate levels associated with low ATP demand. As predicted by molecular dynamic modeling, ANT Lysine23 acetylation decreased the apparent affinity of ADP for ANT binding.

Role of adipocyte mitochondria in inflammation, lipemia and insulin sensitivity in humans: effects of pioglitazone treatment.

BACKGROUND/OBJECTIVES: To gain further insight into the role of adipocyte mitochondria in systemic lipid metabolism, inflammation and insulin sensitivity in humans and to provide a better understanding of the mechanisms of action of the peroxisome proliferator-activated receptor gamma agonist pioglitazone. SUBJECTS/METHODS: Mitochondrial DNA (mtDNA) copy number, mitochondrial distribution, mitochondrial and overall cellular protein abundances as well as intrinsic mitochondrial function of subcutaneous adipocytes were assessed by real-time quantitative PCR, MitoTracker staining, global proteomics analyses and NADH cytochrome c reductase activity in insulin-sensitive, normal-glucose-tolerant (NGT) individuals and age, gender, adiposity-matched insulin-resistant individuals with abnormal glucose tolerant (AGT) before and after 3 months of pioglitazone treatment. RESULTS: mtDNA copy number/adipocyte and mtDNA copy number/adipocyte volume were ~55% and ~4-fold lower in AGT than in NGT, respectively, and correlated positively with the M-value of euglycemic clamps and high-density lipoprotein, and negatively with fasting plasma triglyceride, tumor necrosis factor-α and interleukin-6 levels in the entire cohort. mtDNA copy number/adipocyte volume also correlated positively with plasma adiponectin. Pioglitazone, which improved insulin sensitivity, plasma lipids and inflammation, increased the mitochondrial copy number, and led to a redistribution of mitochondria from a punctate to a more reticular pattern as observed in NGT. This was accompanied by disproportionately increased abundances of mitochondrial proteins, including those involved in fat oxidation and triglyceride synthesis. Pioglitazone also increased the abundance of collagen VI and decreased the abundance of cytoskeletal proteins. NADH cytochrome c reductase activity of isolated adipocyte mitochondria was similar in AGT and NGT and unaltered by pioglitazone. CONCLUSIONS: Adipocyte mitochondria are deficient in insulin-resistant individuals and correlate with systemic lipid metabolism, inflammation and insulin sensitivity. Pioglitazone induces mitochondrial biogenesis and reorganization as well as the synthesis of mitochondrial proteins including those critical for lipid metabolism. It also alters extracellular matrix and cytoskeletal proteins. The intrinsic function of adipocyte mitochondria appears unaffected in insulin resistance and by pioglitazone.International Journal of Obesity advance online publication, 31 October 2017; doi:10.1038/ijo.2017.192.

Haploinsufficiency of protamine-1 or -2 causes infertility in mice.

Protamines are the major DNA-binding proteins in the nucleus of sperm in most vertebrates and package the DNA in a volume less than 5% of a somatic cell nucleus. Many mammals have one protamine, but a few species, including humans and mice, have two. Here we use gene targeting to determine if the second protamine provides redundancy to an essential process, or if both protamines are necessary. We disrupted the coding sequence of one allele of either Prm1 or Prm2 in embryonic stem (ES) cells derived from 129-strain mice, and injected them into blastocysts from C57BL/6-strain mice. Male chimeras produced 129-genotype sperm with disrupted Prm1 or Prm2 alleles, but failed to sire offspring carrying the 129 genome. We also found that a decrease in the amount of either protamine disrupts nuclear formation, processing of protamine-2 and normal sperm function. Our studies show that both protamines are essential and that haploinsufficiency caused by a mutation in one allele of Prm1 or Prm2 prevents genetic transmission of both mutant and wild-type alleles.

MI-63: a novel small-molecule inhibitor targets MDM2 and induces apoptosis in embryonal and alveolar rhabdomyosarcoma cells with wild-type p53.

BACKGROUND: Interruption of the role of p53s as a tumour suppressor by MDM2 may be one of the mechanisms by which cancer cells evade current therapy. Blocking the inhibition of wild-type p53 by MDM2 in cancer cells should reactivate p53's tumour suppressor functions and enhance current cancer treatments. MI-63 is a novel non-peptide small molecule that has shown strong binding affinity (K(i)=3 nM) for MDM2; however, its effects on paediatric cancer cells and the specific mechanism of tumour suppressor reactivation have not been evaluated. METHODS: Rhabdomyosarcoma (RMS), the most common childhood soft tissue sarcoma, expresses either wild-type or mutant p53 protein. We examined the inhibitory effects of MI-63 in embryonal RMS (ERMS) and alveolar RMS (ARMS) cell lines expressing wild-type or mutated p53. RESULTS: Treatment with MI-63 reduced cell viability by 13.4% and by <1%, respectively, at 72 h in both RH36 and RH18 cell lines expressing wild-type p53. In contrast, RH30 and RD2 cells expressing p53 mutants are resistant to MI-63 treatment. An increased expression of p53, p21(WAF1), and Bax protein was observed after treatment with MI-63 in RMS cells with wild-type p53, and apoptosis was confirmed by cleaved PARP and caspase-3 expression. However, RD2 and RH30 RMS cells, as well as human normal skeletal muscle cells, showed a minimal increase in p53 signalling and no induction of cleaved PARP and caspase-3. MI-63 was compared with Nutlin-3, a known MDM2 inhibitor, and was found to be more potent in the inhibition of cell proliferation/viability. Further, synergy was observed when MI-63 was used in combination with doxorubicin. CONCLUSION: These results indicate that MI-63 is a potent therapeutic agent for RMS cells expressing wild-type p53 protein.

Spinothalamic tract neurons in the substantia gelatinosa.

The substantia gelatinosa of the mammalian spinal cord is generally believed to be a closed system; that is its neurons are thought to project only to the substantia gelatinosa of the same or the contralateral side. Experiments in monkeys, using injections of the marker enzyme horseradish peroxidase, show that at least some neurons of the substantia gelatinosa project to the thalamus and thus belong to the spinothalamic tract. Such neurons include two cell types intrinsic to the gelatinosa, the central cells and the limitrophe cells of Cajal.

Nitrification in paleocene shale.

Exchangeable ammonium nitrogen is present in Paleocene (Fort Union) shale below a depth of 10 meters in North Dakota and eastern Montana. Above 10 meters, exchangeable ammonium nitrogen is nitrified in situ. The lack of viable nitrifying organisms and the probable lack of oxygen prevent in situ nitrification below 10 meters. Shale samples incubated at 27 degrees C under nonsterile conditions or shales exposed to atmospheric contamination exhibited active nitrification without additional treatment.

Utilizing fungus myceliated grain for molt induction and performance in commercial laying hens.

Molting in poultry is used to rejuvenate hens for a second or third laying cycle. Feed withdrawal was once the most effective method used for molt induction; however, it has being phased out due to food safety and animal welfare concerns. This study evaluated the utilization of fungus myceliated grain as a safe and effective alternative for inducing molt, enhancing immunity, reducing Salmonella growth, and returning to egg production. Laying hens were subjected to 1 of 5 treatments: 1) nonfed (NF), 2) full-fed (FF), 3) fungus myceliated meal (FM), 4) 90% fungus myceliated meal+10% standard layer ration (FM-90), and 5) 90% alfalfa meal+10% fungus myceliated meal (AF-90). Each treatment condition was replicated 9 times during a 9-d molt period. The results revealed that egg production for treatments 1 and 3 ceased completely by d 5, whereas hens in treatments 4 and 5 ceased egg production by d 6. The percentage of BW loss decreased significantly (P<0.05) in treatments 1 (57%), 2 (8%), 3 (35%), 4 (37%), and 5 (44%). Ovary weights of hens fed all molting diets decreased significantly from the full-fed control but did not differ significantly (P<0.05) from each other. Salmonella population in the crop, ovary, and ceca from hens differed significantly (P<0.05) among treatments. Return to egg production differed between treatments with higher production beginning in treatment 3 and ending in treatment 5. Antibody titers did differ (P<0.05) among treatments. From these results, fungus myceliated meal appears to be a viable alternative to conventional feed withdrawal and other methods for the successful induction of molt and retention of postmolt performance.

Investigating the effects of dietary probiotic feeding regimens on broiler chicken production and Campylobacter jejuni presence.

An experiment was conducted to investigate the effect of restricted feeding of a commercially available probiotic diet on production/processing performance, Campylobacter jejuni prevalence, and organ weights in broiler chickens. Five hundred forty 1-d-old broiler chicks were randomly assigned to a control or a direct-fed microbial (PrimaLac, DFM) diet and subjected to ad libitum full-fed (A), restricted 8-h (R), or skip-a-day (S) feeding regimens. Each of the 6 treatments was replicated 3 times with 15 male and 15 female chicks per pen for 49 d. Significant (P<0.05) differences between BW in the control and DFM groups with regard to feed type were found at d 7 (A), female d 21 (R) male and females, and d 49 (A and S) male and females. Body weights of males in the control group were significantly higher than the DFM (A) and differed by regimens (A>R>S) at d 49, whereas weights of females did not differ in regimens A and S. Body weight in the control females of regimen R was significantly higher than those in regimens A and S. Carcass yield was significantly higher for males in the control regimen A, 78.1 vs. 74.6% for the DFM regimen A; however, females did not differ significantly in this regimen, but did so in regimen S with 72.6 vs. 69.0%. The gizzard weights were significantly higher for broilers exposed to S and R regimens when compared with the A regimen. The prevalence of C. jejuni in the DFM-treated broilers regimen R was lower (33 vs. 60% positive) for the control group at 21 d. The weekly BW throughout the study reflected many variations, but broiler chickens receiving the control feed on regimen A performed better than those receiving the DFM feed. From the present results, it was concluded that supplementation of DFM reduced the presence of C. jejuni but had no significant effect on the growth performance of broilers; however, there were some significant trends regarding sex, feed, and feeding methods on the performance results.

The effect of mushroom and pokeweed extract on salmonella, egg production, and weight loss in molting hens.

An experiment was conducted to evaluate the effects of mushroom and pokeweed extract alone or in combination with alfalfa meal on Salmonella spp. population, egg production, and weight loss in laying hens during a 10-d molting period. The trial used 54 active laying hens approximately 77 wk of age that were naturally infected with Salmonella spp. The layers were subjected to 1 of 9 treatment groups, replicated 3 times with 2 hens per replicate cage. The treatment conditions were as follows: 1) full-fed + H(2)0 (FFW), 2) full-fed + mushroom (FFM), 3) full-fed + pokeweed (FFP), 4) nonfed + H(2)0 (NFW), 5) nonfed + mushroom (NFM), 6) nonfed + pokeweed (NFP), 7) full-fed alfalfa meal + H(2)0 (FFAW), 8) full-fed alfalfa meal + mushroom (FFAM), and 9) full-fed alfalfa meal + poke-weed (FFAP). The results showed that the base-10 logarithm values of Salmonella from the ceca significantly increased (P

Mechanisms of the puromycin-induced defects in the transglomerular passage of water and macromolecules.

To investigate the mechanism(s) of increased filtration of serum proteins after glomerular injury, polydisperse samples of uncharged [(3)H]dextran (D) or anionic [(3)H]dextran sulfate (DS) were infused into 14 control and 16 puromycin aminonucleoside- (PAN) treated Munich-Wistar rats. Fractional clearances of D or DS ranging in radius from 18 to 42A were determined in these rats, together with direct measurements of the forces governing the glomerular filtration rate of water. Whole kidney and single nephron glomerular filtration rates were approximately 40% lower in PAN-treated rats, relative to controls, due mainly to a marked reduction in the glomerular capillary ultrafiltration coefficient and, to a lesser extent, to a small reduction in glomerular plasma flow rate as well. In PAN-treated rats, as in normal controls, inulin was found to permeate the glomerular capillary wall without measurable restriction, and both D and DS were shown to be neither secreted nor reabsorbed. Fractional clearances of uncharged D were reduced after PAN administration, falling significantly for effective D radii from 22 to 38A. Utilizing a theory based on macromolecular transport through pores, these results indicate that in PAN-treated rats, effective pore radius is the same as in controls, approximately 44A. In PAN nephrosis, however, the ratio of total pore surface area/pore length, a measure of pore density, is reduced to approximately one-third that of control, due very likely to a reduction in filtration surface area. In contrast to the results with uncharged D, fractional clearances of DS were found to increase after PAN administration for all DS radii studied. These results with D and DS suggest that proteinuria in PAN nephrosis is due, not to an increase in effective pore radius or number of pores, but rather to a diminution of the electrostatic barrier function of the glomerular capillary wall, thereby allowing increased passage of polyanions such as DS and albumin.

Activation of protein kinase B/Akt in the periphery contributes to pain behavior induced by capsaicin in rats.

Protein kinase B (PKB/Akt) is a member of the second-messenger regulated subfamily of protein kinases. It is implicated in signaling downstream of growth factors, insulin receptor tyrosine kinases and phosphoinositide 3-kinase (PI3K). Current studies indicate that nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and PI3K help mediate inflammatory hyperalgesia. However, little is known about the role of PKB/Akt in the nociceptive system. In this study, we investigated whether PKB/Akt in primary sensory neurons is activated after noxious stimulation and contributes to pain behavior induced in rats by capsaicin. We demonstrated that phospho-PKB/Akt (p-PKB/Akt) is increased in dorsal root ganglia (DRG) at 5 min after intradermal injection of capsaicin. p-PKB/Akt is distributed predominantly in small- and medium-sized DRG cells. After capsaicin injection, p-PKB/Akt (473) is colocalized with isotectin-B4 (IB4), tyrosine kinase A (TrkA), and calcitonin gene-related peptide (CGRP). Furthermore, most transient receptor potential vanilloid type 1 (TRPV1) positive DRG neurons double label for p-PKB/Akt. Behavioral experiments show that intradermal injection of a PI3K (upstream of PKB/Akt) inhibitor, wortmannin, dose-dependently inhibits the changes in exploratory behavior evoked by capsaicin injection. The PKB/Akt inhibitor, Akt inhibitor IV, has the same effect. The results suggest that the PKB/Akt signaling pathway in the periphery is activated by noxious stimulation and contributes to pain behavior.

Performance assessment of broiler chickens given mushroom extract alone or in combination with probiotics.

A study was conducted to evaluate the effect of combined Shiitake mushroom (Lentinus edodes) extract with probiotics (PrimaLac) on the growth and health of broiler chickens. In trial 1, 540 d-of-hatch chicks were randomly assigned to 6 treatment groups, replicated 3 times, with 15 males and 15 females per pen for 3 wk. Dietary probiotics and mushroom treatments were as follows: 1) control feed + ad libitum tap water; 2) control feed + skip-a-day mushroom water; 3) control feed + ad libitum mushroom water; 4) probiotic feed + ad libitum tap water; 5) probiotic feed + skip-a-day mushroom water; 6) probiotic feed + ad libitum mushroom water. Body weight gain, feed consumption and efficiency, mortality, bursa, liver, and spleen relative weights of chicks were taken. In trial 2, the performance of broilers 3 to 7 wk withdrawn from the mushroom extract was evaluated along with the comparative level of fecal biofidobacteria in the control and mushroom extract treatment (trt). Mortality, weight gain, feed consumption and efficiency, carcass yield, fat pads, bursa weights and fecal bifidobacteria were measured in trial 2. In trial 1, significant differences (P < 0.05) in female weight gain (trt 4-0.62 vs. trt 1-0.54 kg) and male spleen weights were observed. In trial 2, significant differences were observed in male weight gain (trt 2-2.40 vs. trt 4-1.12 kg), male and female fat pads, male bursa weights (trt 3-0.15 vs. trt 6-0.39), female carcass yield percentage (trt 1-77.8 vs. trt 4-66.4), and feed consumption and efficiency. Body weights were severely depressed in the male broilers receiving the probiotics feed in treatments 4, 5, and 6, but not in the female broilers. These results indicate that performance differences in gender occur with additives during different grow-out periods, and mushroom extract promotes bifidobacteria growth in broiler chickens after 4 wk of withdrawal. It appears that probiotics and mushroom extract offered no combination potential for weight gain, which was compromised in this study, but possible health-enhanced attributes.

Fiber-type-related differences in the enzymes of a proposed substrate cycle.

A substrate cycle between citric acid cycle (CAC) intermediates isocitrate and 2-oxoglutarate, involving NAD+- and NADP+-linked isocitrate dehydrogenase (NAD-IDH and NADP-IDH, respectively) and mitochondrial transhydrogenase (H+-Thase), has recently been proposed. This cycle has been hypothesized to enhance mitochondrial respiratory control by increasing the sensitivity of NAD-IDH to its modulators and allowing for enhanced increases in flux through this step of the CAC during periods of increased ATP demand. The activities of the enzymes comprising the substrate cycle: NAD-IDH, forward and reverse NADP-IDH, and forward and reverse H+-Thase, along with the activity of a marker of mitochondrial content, citrate synthase (CS) were measured in mitochondria isolated from rabbit Type I and Type IIb muscles and in whole muscle homogenates, representing the various fiber types, from rats. In isolated rabbit muscle mitochondria, NAD-IDH had significantly higher (1.6 x ) activity in white muscle while forward NADP-IDH, forward and reverse H+-Thase, and CS all had significantly higher (1.2-1.6 x ) activities in red muscle. There was no difference in reverse NADP-IDH between fiber types. Similarly, in rat whole muscle enzyme activities normalized to CS, NAD-IDH had significantly higher activity in fast-twitch glycolytic (FG) fibers, while forward NADP-IDH and forward H+-Thase had significantly higher activities in slow-twitch oxidative (SO) fibers. These results suggest that differences in the activities of the substrate cycle enzymes between skeletal muscle fiber types could contribute to differences in respiratory control due to differential cycling rates and/or loci of control.

Neural substrates of childhood attention-deficit/hyperactivity disorder: electroencephalographic and magnetic resonance imaging evidence.

Research methods based on electroencephalogram (EEG) and anatomical and functional MRI have been used with increasing frequency in the study of childhood Attention-Deficit/Hyperactivity Disorder (ADHD). Both methods are safe and noninvasive, and their results can complement each other because of the good temporal (but relatively poorer spatial) resolution of EEG and the good spatial (but relatively poorer temporal) resolution of MRI. These methods are described, and associated recent research on childhood ADHD is summarized and critically examined. Results of this research support theories of ADHD that focus on a frontal-striatal neurological circuitry substrate, which has been implicated in neuropsychological executive functioning. A number of issues, however, such as the specificity of this finding for ADHD, remain unresolved. We conclude with an overview of advances and issues to be considered in future research on the neural substrates of childhood ADHD and advocate a developmental-contextual perspective on this disorder that acknowledges the reciprocal relations between neural structures and functions.

Antagonism of morphine analgesia by nonopioid cold-water swim analgesia: direct evidence for collateral inhibition.

The demonstrated existence of opioid and nonopioid forms of pain control has raised questions as to how they interact. Previous indirect evidence suggests that activation of one system inhibited the activation of the other. The present study assessed this directly using morphine as an opiate form of analgesia and continuous cold-water swims (CCWS, 4 degrees C, 2 min) as the nonopioid form. A significant reduction in morphine (8 mg/kg, SC) analgesia on the tail-flick test was observed if rats were acutely exposed to CCWS immediately prior to morphine administration. The inability of naloxone (10 mg/kg, SC) to reduce CCWS analgesia verified its nonopioid nature. The antagonism of morphine (3 mg/kg, SC) analgesia was greater following preexposure to 2 min of CCWS than 1 min of CCWS. CCWS was also more effective in antagonizing analgesia induced by the 3 mg/kg than the 8 mg/kg dose of morphine. The antagonism of morphine analgesia by CCWS was dependent upon the temporal patterning of stimulus presentation: exposure to CCWS 20 or 60 min prior to morphine failed to alter subsequent morphine analgesia. A significant reduction in analgesia induced by intraperitoneal administration of morphine (10 mg/kg) was also observed when CCWS was presented immediately prior to injection, suggesting that pharmacokinetic factors such as altered drug absorbance by CCWS-induced vasoconstriction do not appear to explain these effects. These data provide direct support for the existence of collateral inhibitory mechanisms activated by CCWS and morphine, and suggests that these opioid and nonopioid forms of analgesia do not function synergistically, but instead involve some form of hierarchical order.

Distribution of the postsynaptic dorsal column projection in the cuneate nucleus of monkeys.

Cells in the spinal cord that are postsynaptic to primary afferent fibers project to the dorsal column nuclei in the postsynaptic dorsal column pathway. The projection of cells in the cervical spinal cord of monkeys to the cuneate nucleus has been reported to avoid pars rotunda of that nucleus, the part that contains the somatotopic representation of the ipsilateral hand. We used the sensitive anterograde tracer Phaseolus vulgaris leucoagglutinin to reexamine this projection. We made multiple iontophoretic injections into the cervical enlargements of three monkeys (two Macaca fascicularis and one Macaca mulatta). Control injections were made in the contralateral dorsal columns of one of these and in the dorsal roots of a fourth animal (M. fascicularis) to test for transport by fibers of passage. After 28-39 days, the animals were deeply anesthetized and perfused, and the tissue was processed for immunohistochemical detection of the label. In all cases (excluding control injections), labeled fibers and varicosities were distributed widely in the ipsilateral cuneate and external cuneate nuclei, including pars rotunda. The dorsal column nuclei ipsilateral to control injections contained no label or only very few poorly labeled fibers, indicating that labeling through fibers of passage did not contribute importantly to the results. This study indicates that the postsynaptic projection to the cuneate nucleus is widespread and includes pars rotunda. Such projections may contribute to transmission of information originating in nociceptors through the dorsal column-medial lemniscal system to the ventrobasal thalamus.

Collaterals of spinothalamic cells in the rat.

Spinothalamic (STT) cells were investigated in the rat to determine the distribution of subpopulations with terminals in both the lateral and medial thalamus, the thalamus bilaterally, or the thalamus and the medullary reticular formation. Two or more retrogradely transported substances (fluorescent dyes, and/or horseradish peroxidase) were injected in each animal. Three combinations of injections were most commonly used: (1) injections of the medullary reticular formation and thalamus, (2) separate injections into each side of the thalamus, and (3) separate injections into the medial and lateral thalamus. The distribution of single labeled cells after each injection was compared with previously published results for rats. The distribution of cells which contained both tracers, double-labeled (DL) cells, was the focus of this study. An average of 15% of STT cells and 8% of spinoreticular cells projected to both the reticular formation and thalamus. However, only a small component of STT cells (less than 2%) projected bilaterally into the thalamus. Most DL cells were found in upper cervical segments. The laminar distribution of all three groups of DL neurons were similar. These cells were most often located in the reticulated part of lamina V and the intermediate zone, lamina VII. STT cells that had terminals in both the medial and lateral thalamus and STT cells with collaterals in the reticular formation were concentrated on the side contralateral to their terminals. These DL neurons provide an anatomical substrate for noxious stimuli to stimuli to activate the reticular formation and thalamus and/or specific sensory and intralaminar thalamus simultaneously.

The organization of the electromotor nucleus and extraocular motor nuclei in the stargazer (Astroscopus y-graecum).

The organization of the oculomotor and electromotor systems was examined in the stargazer, a teleost. The electromotor system in these animals is a derivative of the oculomotor system. The extraocular motor nuclei and nerves consist of approximately equal numbers of motoneurons and axons (about 100 per muscle). In contrast, electromotor axons appear to branch several times within the intracranial portion of the IIIrd nerve. The topographical organization of the motoneurons was examined using retrograde transport of horseradish peroxidase injected into the electric organ or eye muscles. Electromotor and oculomotor neurons form distinct populations. Each electric organ receives a strong ipsilateral and a weak contralateral innervation. Individual eye muscles receive unilateral innervations with the expected laterality. Within the oculomotor nucleus there is some topographical separation of motoneurons innervating each muscle. Antidromic field potentials confirm the identity of the electromotor nucleus.

Glutamate-immunoreactive terminals synapse on primate spinothalamic tract cells.

Glutamate has been shown to excite spinothalamic tract (STT) neurons and has been localized to primary afferent neurons, spinal cord projection neurons, and interneurons in the spinal cord dorsal horn. The likelihood that glutamate-immunoreactive (GLU-IR) terminals directly innervate STT neurons was investigated. For these studies three lamina IV or V STT cells in the lumbar spinal cords of three monkeys (Macaca fascicularis) were identified electrophysiologically and characterized. Two were identified as high threshold neurons and one as a wide dynamic range neuron. Following intracellular injection of the cells with HRP and reaction to give the cells a Golgi-like appearance, the tissues were processed for electron microscopy. Postembedding immunogold methods with antibodies specific for glutamate were used to identify GLU-IR terminals apposing the somata and dendrites of the STT neurons, including dendrites that extended into laminae IV and III. The GLU-IR terminals were numerous and constituted a mean of 46% of the population counted that appose the STT soma and 50% of the profiles apposing the dendrites. Fifty-four percent of the somatic and 50% of the dendritic surface length was contacted by GLU-IR terminals. Most terminals contained round clear vesicles and some contained a variable number of large dense core vesicles. For one of the three cells examined it was determined that 45% of the terminals apposing the soma were GLU-IR and 30% of the terminals were gamma aminobutyric acid-immunoreactive (GABA-IR). In an additional monkey, a lamina I cell retrogradely labeled from the ventral posterolateral nucleus of the thalamus was found to be ensheathed in glial processes.(ABSTRACT TRUNCATED AT 250 WORDS)