Neurokinin A-like immunoreactivity in rat primary sensory neurons; coexistence with substance P

Rat spinal cord, dorsal root ganglia and skin were investigated employing immunohistochemical technique with specific antisera to neurokinin A and substance P. Neurokinin A-like immunoreactivity was detected in the spinal dorsal horn and skin with a similar distribution pattern as that of substance P-like immunoreactivity. After dorsal root transection a parallel decrease of neurokinin A and substance P-like immunoreactivity was observed in the dorsal horn. Using colchicine pretreatment a population of neurokinin A positive cell bodies was seen in the dorsal root ganglia, and by comparison of consecutive sections of the same cells stained for substance P it was revealed that these neurons also display substance P-like immunoreactivity. However, substance P-, but not neurokinin A-, immunoreactive cells were also observed. It is concluded that neurokinin A- and substance P-like immunoreactivity coexist in a population of rat primary sensory neurons.     

Neurokinin A-like immunoreactivity in rat primary sensory neurons; coexistence with substance P

Summary Rat spinal cord, dorsal root ganglia and skin were investigated employing immunohistochemical technique with specific antisera to neurokinin A and substance P. Neurokinin A-like immunoreactivity was detected in the spinal dorsal horn and skin with a similar distribution pattern as that of substance P-like immunoreactivity. After dorsal root transection a parallell decrease of neurokinin A and substance P-like immunoreactivity was observed in the dorsal horn. Using colchicine pretreatment a population of neurokinin A positive cell bodies was seen in the dorsal root ganglia, and by comparison of consecutive sections of the same cells stained for substance P it was revealed that these neurons also display substance P-like immunoreactivity. However, substance P-, but not neurokinin A-, immunoreactive cells were also observed. It is concluded that neurokinin A- and substance P-like immunoreactivity coexist in a population of rat primary sensory neurons.     

Hypothalamic substance P as a candidate for transmitter of primary afferent neurons.

A peptide that exerts a depolarizing action on frog spinal motoneurons was found in the dorsal root of bovine spinal nerve. Pharmacological, chemical, and immunological properties of this motoneuron-depolarizing peptide were investigated and the results indicated that the peptide is identical with an undecapeptide, substance P, recently isolated from bovine hypothalamus by M.M.Chang and S.E.Leeman. The amount of hypothalamic substance P in bovine dorsal root determined by bioassay or radioimmunoassay was 24-130 pmole/g wet wt, whereas that in the ventral root was 9-27 times less. The effects of synthetic hypothalamic substance P on the isolated spinal cord of the frog and the newborn rat were studied. The peptide exerted a powerful depolarizing action on the motoneurons, its potency being about 200 times higher than that of L-glutamate. Distribution of substance P in the cat spinal cord was studied. The concentration of the peptide was highest in the dorsal part of dy lowered. When the dorsal root of the cat was ligated, substance P accumulated in a high concentration on the ganglion side of the ligature. These results, taken together, support the hypothesis that hypothalamic substance P is an excitatory transmitter of primary afferent neurons.     

水稻不同生育期对荧光物——铕吸收运行的研究初报

本文为探索稻田生态系统营养关系定量分析研究方法的环节之一。作者首次采用标记示踪法测定了水稻在秧苗期,分蘖期,壮苞期,抽穗期,乳熟期等5个生育期吸收铕的情况为：同一处理浓度下,水稻根、茎、叶的铕含量在分蘖期最高,抽穗期最低,水稻中铕含量随生育期呈低-高-低-高的变化规律,秧苗期根、茎、叶的铕含量相近且低;分蘖期叶中铕含量比根和茎明显高;其它3个生育期茎与叶中铕含量相近并且比根部含量高。     

家兔脊神经节内肥大细胞与肽能神经关系的观察

探讨脊神经节内肥大细胞与肽能神经的关系,采用常规组织学染色和免疫组织化学方法对家兔脊神经节内肥大细胞和P物质免疫阳性反应进行观察。结果显示：在P物质免疫反应阳性的神经元和神经纤维周围散在着肥大细胞,表明,脊神经节内,肥大细胞与肽能神经存在着组织形态学上的构筑关系。     

Development of an Antiserum to the Midportion of Substance P: Applications for Biochemical and Anatomical Studies of Substance P-Related Peptide Species in CNS Tissues

This report describes the generation and biochemical characterization of a high-affinity antiserum that recognizes an epitope contained in the midportion sequence of substance P, i.e., substance P4-10. Designated A47, this reagent bound a variety of related peptide species containing the substance P4-10 sequence with apparent equipotency. A double radioimmunoassay procedure was developed that utilized A47, in combination with a traditional high-affinity COOH-terminally directed anti-substance P serum, to provide quantification of mature and immature forms of substance P in CNS tissues. Across most rat CNS areas, levels of substance P-like immunoreactivity were consistently 15% higher when monitored by analyses using A47 versus anti-substance P serum. In the dorsal root ganglia, an apparent enhancement in levels of substance P-like immunoreactivity of approximately 40%, when quantified by analyses using A47 versus anti-substance P serum, was observed; this most likely reflected the presence of an active biosynthetic pool of intermediate processing forms of substance P in this tissue. Coordinated HPLC/radioimmunoassay analyses of extracted dorsal root ganglia tissues demonstrated multiple peaks of immunoreactivity corresponding to mature substance P and to several of its precursor forms found in the normal biosynthetic pathway. Of the total recovered HPLC-fractionated immunoreactivities, that corresponding to the putative immediate precursor to substance P, i.e., substance P-glycine, was the predominant peak. In an additional series of HPLC/radioimmunoassay analyses, selective decreases in immunoreactive peaks corresponding to precursor forms of substance P were observed in dorsal root ganglia tissues from rats treated with the neurotoxic agent capsaicin. These results indicated decreased turnover of substance P as a consequence of drug treatment. Finally, initial immunohistochemical analyses employing affinity-purified A47 produced an unusual pattern of labeling characterized by well defined punctate terminal elements within the superficial aspects of the dorsal horn of the spinal cord.     

Heme oxygenase-2 in primary afferent neurons of the guinea-pig

Sensory ganglia (trigeminal, jugular, nodose, cervical and lumbar dorsal root ganglia) of the guineapig were investigated for the presence of a constitutive carbon monoxide-generating enzyme, heme oxygenase-2 (HO-2). A 36-kDa HO-2-immunoreactive protein was identified by Western blotting in protein extracts from dorsal root ganglia and localized by immunohistochemistry to all neuronal perikarya, including both substance P-positive and substance P-negative neurons, in all ganglia investigated. This ubiquitous distribution points to a general requirement for HO-2 in primary afferent neurons rather than to an association with a specific functionally defined subpopulation. Neither the axons of the sensory neurons nor their peripheral terminals in the skin and around visceral arteries were HO-2 immunoreactive. Explants of dorsal root ganglia with crushes placed on the dorsal roots showed accumulation of the neuropeptide, substance P, at the ganglionic side of the crush, but these axons were non-reactive to HO-2, indicating that there is no substantial transport of HO-2 towards the central ending of these sensory neurons. Collectively, the findings suggest that HO-2 exerts it major effects within the sensory ganglia themselves.     

STUDIES ON THE GROWTH HORMONE OF PLANTS : VI. THE DISTRIBUTION OF THE GROWTH SUBSTANCE IN PLANT TISSUES

1. It is shown that when plant tissues are ground with water the growth substance contained therein is inactivated by the oxidizing enzymes. 2. A simple method of extraction is described which enables the quantitative determination of growth substance in such tissues. 3. The amount and distribution of growth substance in the Avena coleoptile is determined by this method, and it is shown that while the substance does not diffuse out from the lower parts of the coleoptile, it is nevertheless present in considerable amounts, the concentration decreasing steadily with the distance from the tip. 4. Growth substance is also present in considerable amounts in Avena roots, and here also its concentration decreases steadily with distance from the tip. 5. The amount of growth substance diffusing out of root tips into dextrose agar, even during long periods of time, is not greater than the amount obtainable by direct extraction. Actual production in the root tip therefore either does not take place at all, or else takes place under quite different conditions from the production in the tip of the coleoptile.     

Immunocytochemical localization of neuromedin K (neurokinin B) in rat spinal ganglia and cord.

Specific antisera directed against substance P and neuromedin K (neurokinin B) have been used in double-label immunofluorescence studies to unambiguously localize these two neuropeptides of the tachykinin family in single tissue sections of rat spinal cord and dorsal root ganglia. Substance P-like immunoreactivity (SPLI) is present but neuromedin K-like immunoreactivity (NMKLI) is undetectable in dorsal root ganglia. Both peptides are present in the spinal cord, but NMKLI is largely restricted to the dorsal gray while SPLI shows a broader distribution. In the spinal gray, NMKLI coexists with SPLI in some, but not all, fibers. While substance P in the dorsal spinal cord is largely of primary afferent origin, neuromedin K appears to originate largely from intrinsic spinal neurons.     

Organ-specific-active allelopathic substance in red pine needles

An organ-specific-growth inhibitory substance was isolated from an aqueous methanol extract of red pine needles and determined by spectral data as 1-mono(16-hydroxyhexadecanoyl)glycerol. This substance inhibited root growth of cress (Lepidium sativum L.) and barnyard grass (Echinochloa crus-galli (L.) Beauv) seedlings at concentrations greater than 0.01 and 0.03???M, respectively. The concentrations required for 50?% growth inhibition on roots of cress and barnyard grass were 0.16 and 3.4???M, respectively. However, the inhibitory activity of the substance on shoots of cress and barnyard grass was very weak. The endogenous concentration of 1-mono(16-hydroxyhexadecanoyl)glycerol in the pine needles was 4.6???mol?kg^?1. Two related compounds, 1-monohexadecanoylglycerol and 16-hydroxyhexadecanlic acid had no activity up to 1,000???M on cress roots and shoots. The effectiveness of 1-mono(16-hydroxyhexadecanoyl)glycerol on root growth inhibition and the occurrence of 1-mono(16-hydroxyhexadecanoyl)glycerol in pine needles suggest the substance may play an important role in the allelopathy of red pine. Root-specific-growth-inhibition by the substance may be one of the strategies for red pine to compete with neighboring plants for nutrients and space because root growth of competitive plants may be very important for their whole plant development.     

Calcitonin gene-related peptide coexists with substance P in capsaicin sensitive neurons and sensory ganglia of the rat

Immunohistochemical and radioimmunoassay studies revealed that both CGRP- and SP-like immunoreactivity in the caudal spinal trigeminal nucleus and tract, the substantia gelatinosa and the dorsal cervical spinal cord as well as in cell bodies of the trigeminal ganglion and the spinal dorsal root ganglion is markedly depleted by capsaicin which is known to cause degeneration of a certain number of primary sensory neurons. Higher brain areas and the ventral spinal cord were not affected by capsaicin treatment. Furthermore CGRP and substance P-like immunoreactivity were shown to be colocalized in the above areas and to coexist in cell bodies of the trigeminal ganglion and the spinal dorsal root ganglia. It is suggested that CGRP, like substance P, may have a neuromodulatory role on nociception and peripheral cardiovascular reflexes.     

Cytokinin Production by Rhizobium japonicum

Possible hormonal interactions between soybean roots and the Rhizobium initiating nodule proliferation in this genus were studied. A cytokinin has been isolated by column and paper chromatography from an effective strain of Rhizobium japonicum grown in pure culture. The substance promotes cell proliferation in a cytokinin-requiring soybean callus tissue. The bacteria are capable of conditioning a cylokinin-free soybean culture medium so that it is able to support the cytokinin-requiring tissue. It is concluded that the substance is a product of the bacteria rather than an artifact of purification. This unidentified cytokinin or a substance moving to a similar Rf value on the paper chromalogram produces polyploid divisions when tested on cultured pea root segments. Some of the division figures exhibit the diploclirornosomes typical of the root nodule primordium in pea.     

Effects of Light on the Georeaction and Growth Inhibitor Content of Roots

The positive geotropic response of the apical segments prepared from the primary roots of Zea mays depends upon at least one growth inhibitor, produced by the root cap, moving basipetally into the extending zone of the root in which it accumulates in the lower part. Anjou maize reacts in both darkness and light while Kelvedon maize is, for the first few hours, geotropic only in light. The production (or activity) of the growth-inhibiting substance — tested by using vertical half-decapitated root segments — is quite similar to the georeaction. This finding provides strong evidence that, in the case of Kelvedon maize roots, the inhibitory substance may depend on light. Observations related to the root segment of Anjou and Kelvedon maizes of which the tips are exchanged, are in agreement with the above results.     

Identification of tryptophan in leachate of oat hulls (Avena sativa) as a mediator of root growth regulation

Leachate of oat hulls ( Avena sativa L. cv. Sang) inhibits root elongation and causes swollen roots and abundant root hair formation. The active substance was isolated by column chromatography and thin layer chromatography (TLC) systems. High performance liquid chromatography (HPLC) revealed 3 peaks, one of which corresponded to the substance responsible. The latter was identified as tryptophan by means of its UV spectrum, amino acid analysis, nuclear magnetic resonance spectrometry (NMR) and mass spectrometry (MS).     

Nerve Growth Factor-like Immunoreactive Substance in Panax ginseng Extract

Panax ginseng extract contains nerve growth factor-like immunoreactive substance (Panax ginseng NGF) that is cross-reactive with anti-mouse NGF IgG. Panax ginseng NGF-like substance and mouse NGF are almost equivalent with respect to their neurite outgrowth-stimulating activity and are immunologically indistinguishable. The molecular weight of Panax ginseng NGF-like substance estimated by the gel filtration method is identical with that of mouse NGF. The isoelectric point of Panax ginseng NGF is about 9.1, like that of mouse NGF. These results suggest that the root of Panax ginseng contains a biologically active NGF-like immunoreactive substance.     

Distribution of five peptides, three general neuroendocrine markers, and two synaptic-vesicle-associated proteins in the spinal cord and dorsal root ganglia of the adult and newborn dog: an immunocytochemical study

This study describes the immunocytochemical distribution of five neuropeptides (calcitonin gene-related peptide [CGRP], enkephalin, galanin, somatostatin, and substance P), three neuronal markers (neurofilament triplet proteins, neuron-specific enolase [NSE], and protein gene product 9.5), and two synaptic-vesicle-associated proteins (synapsin I and synaptophysin) in the spinal cord and dorsal root ganglia of adult and newborn dogs. CGRP and substance P were the only peptides detectable at birth in the spinal cord; they were present within a small number of immunoreactive fibers concentrated in laminae I-II. CGRP immunoreactivity was also observed in motoneurons and in dorsal root ganglion cells. In adult animals, all peptides under study were localized to varicose fibers forming rich plexuses within laminae I-III and, to a lesser extent, lamina X and the intermediolateral cell columns. Some dorsal root ganglion neurons were CGRP- and/or substance P-immunoreactive. The other antigens were present in the spinal cord and dorsal root ganglia of both adult and newborn animals, with the exception of NSE, which, at birth, was not detectable in spinal cord neurons. Moreover, synapsin I/synaptophysin immunoreactivity, at birth, was restricted to laminae I-II, while in adult dogs, immunostaining was observed in terminal-like elements throughout the spinal neuropil. These results suggest that in the dog spinal cord and dorsal root ganglia, peptide-containing pathways complete their development during postnatal life, together with the full expression of NSE and synapsin I/synaptophysin immunoreactivities. In adulthood, peptide distribution is similar to that described in other mammals, although a relative absence of immunoreactive cell bodies was observed in the spinal cord.     

Excitatory responses of trigeminal neurons to substance P suggest involvement in sensory transmission

Responses to substance P application were studied with intracellular recording techniques in in vitro preparations of trigeminal root ganglion neurons of guinea pigs. Perfusion of substance P in micromolar concentrations markedly depolarized neurons and reduced their input conductances. Also, the threshold for spikes evoked by injections of depolarizing current pulses was decreased. Single electrode voltage-clamp recordings showed that substance P increased inward, and decreased outward currents evoked by hyperpolarizing voltage steps from holding potentials near rest. Depolarizing responses to substance P were attenuated in Na+-deficient solutions. The excitatory actions of this endogenous peptide on the perikarya of primary sensory neurons give rise to the possibility of physiological actions of substance P at multiple sites in the trigeminal system.     

Cytokinin Production by Tomato Root: Occurrence of Cytokinins in Staled Medium of Root Culture

Cytokinins were released into the medium by cultures of excised tomato (Lycopersicon esculentum Mill.) root tips. A large portion was directly released from the proximal cut-end of the roots. The most active substance was found to co-chromatograph with zeatin riboside in both paper and Sephadex column chromatographies. The cytokinin activity after 7 days of growth was found about eight times higher in the medium as compared with that in the root tissues. When freshly excised root tips were subcultured throughout eight passages, the accumulation of cytokinin in the medium of each passage was approximately the same. These results show that the root tip is a main site of cytokinin synthesis.     

Distribution of L-DOPA in the root of velvet bean plant (Mucuna pruriens L.) and gravity.

Velvet bean (Mucuna pruriens L.) has been found that the degree of suppression on the lettuce root growth by velvet bean was less on the 3D-clinorotation. The number and growth of adventitious root in velvet bean differed among the clinostated and control group. L-DOPA (L-3,4-dihydroxyphenylalanine) is known to be the major substance in the allelopathy of velvet bean plant, released from its root. Since L-DOPA is a precursor of melanin pigment, and is easily converted to melanin by oxidation, locality of L-DOPA production in the plant body can be seen through pigmentation. The amount of L-DOPA was analyzed by HPLC and LC-ESI/MS. The distribution of L-DOPA in the root was different among the ground control condition and pseudo-microgravity.     

水培对羽裂蔓绿绒解剖结构的影响

采用石蜡切片法,对天南星科植物羽裂蔓绿绒(Philodendron pitfieri Engl.)水培和土培根的解剖特征进行观察,探讨蔓绿绒适应水生环境的形态学特点,并比较了水培根与土培根的形态解剖学差异.结果表明:水培后根冠不明显,淀粉体较少,多糖类物质较少而土培根根冠体积较大,淀粉体较多,多糖类物质多,土培根PAS反应强烈;水培根表皮厚度比土培根小,水培根根毛少或退化消失;水培根皮层薄壁组织发达,细胞大型,壁薄,细胞间隙中含有溶生性通气组织,土培根皮层未见通气组织.