Lectins are glycoproteins that specifically bind carbohydrate structures and may participate in the biodefense mechanisms of fish. In this study, the binding of three lectins, Dolichos biflorus agglutinin (DBA), soybean agglutinin (SBA), Bandeiraea simplicifolia BS-1 (isolectin B4), Triticum vulgaris (WGA), Arachis hypogaea (PNA) and Ulex europaeus (UEA-I) were studied in the gill, liver, intestine, kidney, heart, and spleen of the flat fish Paralichthys olivaceus. DBA was detected in intestinal mucous cells, as well as in gill epithelial and mucous cells. It was weakly detected in renal tubule epithelial cells and in bile duct epithelial cells. The strong SBA staining was seen in the intestinal club cells, in bile duct epithelial cells and renal tubule epithelial cells. There were intense positive reactions for isolectin B4 in gill epithelial and mucous cells, and the strong isolectin B4 staining was seen in epithelial cells of the bile duct and intestine. The strong WGA staining was seen in the gill mucosal cells, sinusoid, renal tubule epithelial cells and mucosal cells of the intestine. UEA-I was detected in the gill epithelial and mucosal cells, bile duct epithelial cells and renal tubular epithelial cells. These results suggest that the six lectins examined were localized in the covering epithelia of the various organs of the flat fish and they may participate in the biodefense mechanism of the intra body surface in which is exposed to various antigens.
Animals ; Epithelial Cells/metabolism ; Flatfishes/*metabolism ; Histocytochemistry/veterinary ; Lectins/*metabolism ; Mucus/metabolism ; Peanut Agglutinin/metabolism ; Plant Lectins/metabolism ; Soybean Proteins/metabolism ; Wheat Germ Agglutinins/metabolism

In this study, wheat germ agglutinin (WGA), tomato lectin (TL) and asparagus pea lectin (AL) were covalently coupled to conventional poly lactic-co-glycolic acid (PLGA) nanoparticles using a carbodiimide method to take the bioadhesive properties. The influences of the amounts of activating agents and lectins, as well as the activating time and incubating time on the effect of lectin conjugating were investigated to optimize the preparation conditions. The mean diameters of the performed nanoparticles with or without lectin conjugation ranged from (140.7 +/- 5.7) nm to (245.6 +/- 18.3) nm. The yields of lectin conjugating and the lectin surface concentrations on nanoparticles were determined by Lowry's methods, and were calculated to be (18.97 +/- 2.9)% - (20.15 +/- 2.4)% and (9.46 +/- 1.45)--(10.05 +/- 1.19) microg x mg(-1), respectively. The in vitro bioadhesive activities of nanoparticles were evaluated by pig gastric mucin (PM) binding experiments. After incubation at room temperature for 60 min, the equilibria of binding between nanoparticles and PM reached. The percentages of the bulk PM which had interacted with different lectin-conjugated PLGA nanoparticles were 15.5%, 12.1% and 11.8%, respectively. The conjugation of lectin enhanced the interaction about 2.4 - 3.2 fold compared with that of the non-conjugated one. A mathematical model was used based on the Langmuir equation, and the rate constants of interaction (k) were calculated to be 2.373 x 10(-3), 1.536 x 10(-3) and 1.714 x 10(-3) (microg x min/mL)(-1), respectively. These interactions could be competitively inhibited by their corresponding sugars of lectins. The results suggested that lectin-conjugated PLGA nanoparticles greatly promoted the interaction with PM in vitro compared with the conventional PLGA nanoparticles, thus would improve the bioadhesion on gastrointestinal mucosa after oral administration resulting in a prolonged residence time in the gastrointestinal tract.
Adhesiveness ; Drug Carriers ; Drug Compounding ; Drug Delivery Systems ; Gastric Mucins ; metabolism ; Lactic Acid ; chemistry ; metabolism ; Nanoparticles ; Plant Lectins ; chemistry ; metabolism ; Polyglycolic Acid ; chemistry ; metabolism ; Protein Binding ; Wheat Germ Agglutinins ; chemistry ; metabolism

Previous studies have shown that inhibitory synaptic inputs are different between in spinal and trigeminal motor systems and activities of jaw closing and opening alpha motoneurons are different during a chewing cycle. This study examined the distribution of inhibitory synapses made on masseter and digastric motoneurons by using retrograde tracing of wheat germ agglutinin conjugated to horseradish peroxides (WGA-HRP) combined with postembedding immunogold labeling on serial ultrathin sections.Many boutons IR (immunoreactive) to GABA and/or glycine were found to appose on two kinds of motoneurons, which were containing pleomorphic vesicles (a mixture of round, oval and flattened vesicles) and exhibited symmetrical synaptic contacts on the somata. Packing density and synaptic covering % were higher in digastric than in masseter motoneurons. Of 703 boutons apposing on 12 masseter motoneurons, 6.08+/-3.51, 29.67+/-8.89 and 17.78+/-5.22% were IR to GABA only, glycine only, and both GABA and glycine, respectively. Of 637 boutons apposing on 11 digastric motoneurons, 6.37+/-4.64, 19.74+/-8.25 and 12.01+/-5.38% were IR to GABA only, glycine only, and both GABA and glycine, respectively. Proportions of glycine IR boutons were higher than that of GABA IR boutons in both masseter and digastric motoneurons. Packing density and proportion of boutons IR to GABA and/or glycine were higher in jaw closing than in jaw opening motoneurons (packing density, 12.03+/-1.58 vs 10.28+/-2.99; proportion of IR boutons, 53.54+/-8.94% vs 38.12+/-9.38% in jaw closing and opening motoneurons, respectively). These results provide ultrastructural evidence that GABA and glycine act as important neurotransmitters for modulation of jaw movement and that proportion of inhibitory synapses is higher in jaw closing than in jaw opening motoneurons.
Animals ; Armoracia ; gamma-Aminobutyric Acid ; Glycine ; Jaw ; Mastication ; Neurotransmitter Agents ; Peroxides ; Rats* ; Synapses ; Triticum ; Wheat Germ Agglutinin-Horseradish Peroxidase Conjugate

The distribution of GABA and/or glycine like immunoreactive nerve terminals on the soma of the masseteric gamma motoneurons were investigated using retrograde tracing of WGA-HRP (wheat germ agglutinin conjugated horseradish peroxidase) and postembedding immunogold labeling methods in serial ultrathin sections. Quantitative analysis of 140 nerve terminals apposing on somata of gamma motoneuron size less than 21 mm in average diameter was performed. The results obtained were as follows. 1. Synaptic covering % of apposing nerve terminals was 21.45+/-11.48% and packing density was 12.85+/-6.17. 2. Nerve terminals immunoreactive (IR) to GABA or glycine were F type containing pleomorphic vesicles with round shape predominant. Majority of nerve terminals immunonegative to GABA or glycine were S type containing spherical vesicles and few of them were F type. 3. 11.42+/-10.00% of examined nerve terminals were IR to GABA only, and 12.71+/-9.85% were IR to GABA and glycine, and 15.21+/-9.58% were IR to glycine only. 4. Synaptic covering % of nerve terminals IR to glycine only was highest (4.58+/-4.50%), followed in order by GABA and glycine (3.18+/-2.77%), and GABA only (2.38+/-2.06%). 5. Among all terminals, immunonegative nerve terminals (60.66+/-14.65%) were much more than nerve terminals immunoreactive to GABA and/or glycine (39.34+/-14.65%) These results show that inhibitory synaptic input and synaptic organization of the masseteric gamma motoneurons reveal characteristic features in contrast to that of alpha motoneurons and which may correlated to the electrophysi-ological characteristics of masseteric gamma motoneurons.
Animals ; Armoracia ; Carisoprodol ; gamma-Aminobutyric Acid* ; Glycine* ; Jaw* ; Presynaptic Terminals* ; Rats* ; Wheat Germ Agglutinin-Horseradish Peroxidase Conjugate

Viscum plants,the evergreen perennial parasitic shrubs or subshrubs,are mainly distributed in tropical and subtropical regions. There are about 70 Viscum species around the world,including 11 species and one variety in China. Mistletoe lectins are typeⅡ ribosome-inactivating proteins( RIPs) extracted from Viscum plants with anticancer and immunoregulatory activities. Many studies have focused on the mistletoe lectins isolated from V. album in Europe and V. album var. coloratum distributed in South Korea,respectively,and several preparations,such as Iscucin Ⓡ,were developed and clinically applied for cancer treatment. Although Viscum plants are widely distributed in China,only a few studies of mistletoe lectins have been reported. The recent progress of mistletoe lectins was reviewed from extraction,purification,quantitative/qualitative detection,molecular structure,pharmacological activities,toxicities,and clinical application,aiming at providing a reference for in-depth research and utilization of mistletoe lectins produced in China.
Humans ; Lectins ; Plant Extracts ; Plant Lectins ; Plant Proteins/genetics* ; Toxins, Biological ; Viscum

Histochemical patterns of lectin binding during development of the rat vomeronasal organ (VNO) were studied to determine whether glycoconjugates are differently expressed after birth. Three types of lectins, Dolichos biflorus agglutinin (DBA), wheat germ agglutinin (WGA), and Ulex europaeus agglutinin I (UEA-I), were studied histochemically in the rat VNO at various stages post-birth: postnatal days 1 and 7, the preweaning period (4 weeks after birth), and at sexual maturity (8 weeks after birth). The free border of the vomeronasal sensory epithelium was positive for both WGA and UEA-I in rats of all ages; whereas, VNO receptor cells and supporting cells were positive only for both WGA and UEA-I from 4 weeks after birth. DBA reactivity was detected in the free border but less so in receptor cells and supporting cells. WGA and UEA-I, but not DBA, showed similar patterns in various ages. In the Jacobson's gland, WGA, UEA-I and DBA were detected in some acini from 4 weeks after birth but not at postnatal days 1 or 7. Collectively, reactivity for three lectins, WGA, UEA-I and DBA, increased in receptor cells and gland acini during postnatal development, possibly contributing to the enhanced chemoreception in rats.
Animals ; Dolichos ; Epithelium ; Glycoconjugates ; Lectins ; Parturition ; Plant Lectins ; Rats ; Triticum ; Ulex ; Vomeronasal Organ

The origin of sympathetic and sensory nerves innervating heart in the cat was investigated using HRP (Horseradish peroxidase) and WGA-HRP (Wheat germ agglutinin-horseradish peroxidase) as neuronal tracers. The neural tracers were injected into subepicardial layer and myocardium of the right atrium, left atrium, right ventricle and left ventricle, respectively. Labeled sympathetic neuronal cell bodies were found in superior cervical ganglia, middle cervical ganglia, stellate ganglia and 4th and 5th thoracic ganglia, mainly in middle cervical ganglia and stellate ganglia. Heavier labeled neuronal cell bodies were found in the middle cervical ganglia and stellate ganglia when the neural tracers were injected into left atrium, right ventricle and left ventricle. Labeled sensory neuronal cell bodies were found in nodose ganglia and T1-T6 spinal ganglia, mainly in T1-T5 spinal ganglia. Heavier labeled neuronal cell bodies were found in the nodose ganglia when the neural tracers were injected into left atrium and right ventricle. These results may provide a neuroanatomical data on origin of sensory nerves innervating the heart of the cat.
Animals ; Cats* ; Ganglia ; Ganglia, Sensory ; Ganglia, Spinal ; Ganglia, Sympathetic ; Heart Atria ; Heart Ventricles ; Heart* ; Horseradish Peroxidase ; Myocardium ; Neurons* ; Nodose Ganglion ; Sensory Receptor Cells ; Stellate Ganglion ; Superior Cervical Ganglion ; Wheat Germ Agglutinin-Horseradish Peroxidase Conjugate*

The present study was carried out to investigate the glycoconjugate properties of the nasal mucosa in the rat after inhalation of formaldehyde. Sprague-Dawley male rats were inhalated 30 ppm formaldehyde for 3 times with 3 hours exposure. The olfactory and respiratory mucosa in the nasal mucosa were taken from the animals on 3, 6,9 days and 2, 3, 4, 5 weeks after inhalation of formaldehyde. The properties of glycoconjugate of the olfactory and respiratory mucosa were investigated using nine biotinylated lectins (PSA, UEA I, PHA-L, BSL I, PNA, MAL I, DBA, BSL II or sWGA). In experimental groups, the degenerative changes of the olfactory epithelium were observed until 3 weeks after inhalation of formaldehyde, but the respiratory epithelium was no change. In control group, the olfactory cells in the olfactory epithelium reacted with PSA, UEA I, PNA, DBA, BSL II, sWGA, and the supporting cells reacted with PSA, PHA-L, PNA, MAL I, DBA, BSL II, sWGA, and Bowman's glands reacted with all the lectins. In experimental groups, the olfactory cells reacted with UEA I, DBA, and the supporting cells reacted with PHA-L, MAL I, DBA, UEA I, and the positive reaction of Bowman's glands was increased. In control group, the goblet cells in the respiratory epithelium reacted with UEA I, MAL I, and the ciliated columnar cells reacted with PSA, UEA I, PHA-L, BSL I, DBA, BSL II, sWGA, and the septal nasal glands reacted with all the lectins except UEA I. In experimental groups, the goblet cells reacted with UEA I, MAL I and PNA. Conclusively, the olfactory mucosa was shown a lot of changes in the properties of glycoconjugates following inhalation of formaldehyde, but respiratory mucosa was shown feeble change. These results suggest that there were different sugar residues of glycoconjugate in the olfactory and respiratory mucosa following inhalation of formaldehyde, respectively.
Animals ; Formaldehyde ; Glycoconjugates ; Goblet Cells ; Humans ; Inhalation ; Lectins ; Male ; Nasal Mucosa ; Olfactory Mucosa ; Phytohemagglutinins ; Rats ; Respiratory Mucosa ; Wheat Germ Agglutinins

Neuropathy is a general term referring to disorders of nerves, and produces when the nerves are damaged. It is characterized by spontaneous pain, allodynia and hyperalgesia. The purpose of present study is to observe the number of WGA-HRP (wheat germ agglutinin-horseradish peroxidase) labelded sensory neurons of DRG (dorsal root ganglia), and distributions according to cell size of sensory neuron in tibial nerve ligation model (NLM). The tibial nerve ligation was performed with 3-0 silk by the application of three tight ligatures at the mid-thigh level. In the neuropathy model of rat tibial nerve ligation, morphological changes of sensory neurons in DRG were observed using WGA-HRP. Rats of NLM showed the neuropathic behaviors. Rats were shown guarding affected limb and limping. Their toes and ankle joint of operated limb were hyperflexed. Under light microscopy, tibial nerve showed degeneration of axons in NLM. In control and NLM, labeled sensory neurons of tibial nerve distributed L4 and L5 DRG. In control group, the labeled sensory neurons were round or oval in shape. They were large and small cells, and mixed pattern. Total number of labeled sensory neurons in NLM decreased significantly from control group. The number of labeled sensory neurons in L4 and L5 DRG decreased significantly from control group. Labeled large and small cells decreased significantly from control group. Present study may serve as the basic information about the changes of DRG sensory neurons in NLM.
Animals ; Ankle Joint ; Axons ; Cell Size ; Diagnosis-Related Groups ; Extremities ; Hyperalgesia ; Ligation ; Light ; Microscopy ; Peripheral Nervous System Diseases ; Rats ; Sensory Receptor Cells ; Silk ; Tibial Nerve ; Toes ; Wheat Germ Agglutinin-Horseradish Peroxidase Conjugate

The spicoreticulocerebellar (SRC) tract is an indirect spinocerebellar tract formed by the reticular formation (RF), which is connected to the cerebellum and spinal cord. The RF receives ascending fibers to both the spinal enlargement and sends descending fibers to the cerebellum. This study demonstrated that the connectivity of the neurons in the RF is concerned to the cerebellum and spinal cord using the anterograde projection with biotinylated dextran amine (BDA) and retrograde labeling with wheat germ agglutinin-horseradish peroxidase (WGA-HRP). Until now, a preliminary study in mammals has dealt with the afferent and efferent pathways in separating groups of neurons in the RF. There are only few reports on chickens. This study examined the SRC tract in chickens. Following bilateral injections we injected BDA into chicken spinal cord (lumbosacral enlargement) and WGA-HRP into the cerebellum. Both of single- and double-labeled cells were found within the RF. The spinoreticular axons were mainly distributed from the potomedullary junction to the rostral medulla in the rostro-caudally RF levels, for example, nucleus of reticularis (n. r.) pontis oralis, locus coeruleus, n. r. pontis caudalis, n. r. pars gigantocellularis, n. r. gigantocellularis and n. r. parvocellualris. Reticulocerebellar labeling by the WGA- HRP was found in the same place as well as that of the BDA-projection. We observed that the proportion and location of double labeling cells in the chicken were almost similar in each level, comparing to the rodents. These results suggest that the reticular formation is strongly related to the spicoreticulocerebellar tract in chickens.
Afferent Pathways/physiology ; Animals ; Biotin/*analogs&derivatives ; Cerebellum/anatomy&histology/*physiology ; Chickens/*anatomy&histology/*physiology ; Dextrans ; Efferent Pathways/physiology ; Microinjections ; Reticular Formation/anatomy&histology/*physiology ; Spinal Cord/anatomy&histology/*physiology ; Wheat Germ Agglutinin-Horseradish Peroxidase Conjugate