scholarly journals Immunohistochemical characterization of neurons in the vestibular ganglion (scarpa’s ganglion) of the pig

2012 ◽  
Vol 15 (3) ◽  
pp. 499-507 ◽  
Author(s):  
A. Dudek ◽  
W. Sienkiewicz ◽  
J. Kaleczyc
Keyword(s):  
Nerve Fibers ◽  
Double Labelling ◽  
Immunofluorescence Method ◽  
Double Labeling ◽  
Round Nucleus ◽  
Scarpa's Ganglion ◽  
M 12 ◽  
M 31 ◽  
Labelling Immunofluorescence

Abstract The study was carried out on three 4-month old female pigs. All the animals were deeply anesthetized and transcardially perfused with 4% buffered paraformaldehyde (pH 7.4). Vestibular ganglia (VG) were collected and processed for double-labelling immunofluorescence method. The preparations were examined under the Zeiss LSM 710 confocal microscope equipped with adequate filter blocks. Neurons forming VG were round or oval in shape with a round nucleus in the center. The majority of them (58%) were medium (M) (31-50 μm in diameter) while 28 % and 14% were small (S) (up to 30 μm in diameter) or large (L) (above 50 μm in diameter) in size, respectively. Double-labeling immunofluorescence revealed that VG neurons stained for CGRP (approx. 81%; among them 70.5%, 26.2% and 3.3% were M, S and L in size, respectively), VACHT (57%; 63% M, 24% S, 13% L), Met-Enk (25%; 60% M, 12% S, 28% L), VIP (20%; 88% M, 6% S, L), NPY (15%; 67% M, 20% S, 13% L), GAL (15%; 74% M, 21% S, 5% L), SP (12%; 69% M, 25% S, 6% L) and NOS-positive (12%; 50% S, 50% M). The most abundant populations of intraganglionic nerve fibers were those which stained for CGRP or Met-Enk, whereas only single SP- or NOS-positive nerve terminals were observed.

Immunohistochemical characterization of the jugular (superior vagal) ganglion in the pig

2017 ◽  
Vol 20 (2) ◽  
pp. 377-385 ◽  
Author(s):  
W. Sienkiewicz ◽  
A. Dudek ◽  
A. Zacharko-Siembida ◽  
M. Marszałek
Keyword(s):  
Nerve Fibers ◽  
Double Labeling ◽  
Round Nucleus ◽  
M 12 ◽  
M 31 ◽  
Left And Right ◽  
Immunofluorescence Labeling ◽  
Labeling Method ◽  
Vagal Ganglia

Abstract The study was carried out on three 4-month old female pigs. All the animals were deeply anesthetized and transcardially perfused with 4% buffered paraformaldehyde (pH 7.4). Left and right superior vagal ganglia (SVG) were collected and processed for immunofluorescence labeling method. The preparations were examined under a Zeiss LSM 710 confocal microscope equipped with adequate filter block. Neurons forming SVG were round or oval in shape with a round nucleus in the center. The majority of them (52%) were medium (M) (31-50 μm in diameter) while 7% and 41% were small (S) (up to 30μm in diameter) or large (L) (above 50 μm in diameter) in size, respectively. Double-labeling immunofluorescence revealed that SVG neurons stained for CGRP (approx. 57%; among them 37%, 9% and 54% were M, S and L in size, respectively), SP (14.5%; 72.4% M, 3.4% S, 24.2% L), VACHT (26%; 63% M, 24% S and 13% L), GAL (14%; 57% M, 29% S, 14% L), NPY (12%; 53% M, 12% S, 35% L), Met-Enk (5%; 40% M, 6% S and 54% L), PACAP (15%; 52% M, 24% S and 24% L), VIP (6.3%; 67% M, 8% S and 25% L), and NOS-positive (6%; 31% M and 69% L). The most abundant populations of intraganglionic nerve fibers were those which stained for CGRP or GAL, whereas only single SP-, PACAP- or Met-ENK-positive nerve terminals were observed.

scholarly journals Immunohistochemical characterization of efferent neurons innervating the oviduct in the pig located in the sympathetic chain ganglia

2012 ◽  
Vol 47 (No. 4) ◽  
pp. 85-91
Author(s):  
K. Czaja
Keyword(s):  
Nerve Cells ◽  
Sympathetic Chain ◽  
Double Labelling ◽  
Retrograde Labelling ◽  
Calcitonin Gene ◽  
Ovum Transport ◽  
Efferent Neurons ◽  
Oxide Synthase ◽  
Labelling Immunofluorescence

The present study was aimed at disclosing the pattern(s) of putative co-incidence of tyrosine hydroxylase (TH), dopamine -hydroxylase (DH), neuropeptide Y (NPY), substance P (SP), calcitonin gene-related peptide (CGRP) and nitric oxide synthase (NOS) within the porcine &ldquo;oviductal&rdquo; efferent neurons using combined retrograde tracing and double-labelling immunohistochemistry. The fluorescent retrograde tracer Fast Blue (FB) was injected into the wall of the ampullar (n = 5) and isthmal (n = 5) part of the organ in ten sexually immature female pigs. After a survival period of three weeks sympathetic chain ganglia (SCG) were collected. 10 &micro;m-thick cryostat sections of the ganglia were examined for the presence of FB-positive (FB<sup>+</sup>) nerve cells under the fluorescent microscope. Tracered neurons were processed for double-labelling immunofluorescence according to the method of Wessendorf and Elde. Retrograde labelling revealed a population of &ldquo;oviductal&rdquo; efferent neurons located in the thoracic (T) and lumbar (L) SCG at the level of T<sub>14</sub> to L<sub>5</sub>. Double-labelling immunofluorescence allowed several subpopulations of the studied perikarya to be distinguished. The largest one consisted of TH<sup>+</sup>/DH<sup>+</sup> (immunopositive) nerve cells. The moderate number of FB<sup>+</sup> nerve cells expressed TH/NPY- immunoreactivity (IR). The tracered neurons did not show SP, CGRP and NOS immunoreactivity. Because identically coded nerve fibres have been observed within the wall of the porcine oviduct it can be assumed that TH<sup>+</sup>/DH<sup>+</sup> or TH<sup>+</sup>/NPY<sup>+</sup> neurons are involved in the control the oviductal tonus and ovum transport.

scholarly journals Characterization of an immunofluorescence technique for the demonstration of coexisting neurotransmitters within nerve fibers and terminals.

1985 ◽  
Vol 33 (10) ◽  
pp. 984-994 ◽  
Author(s):  
M W Wessendorf ◽  
R P Elde
Keyword(s):  
Substance P ◽  
Central Canal ◽  
Cross Reactivity ◽  
Immunohistochemical Localization ◽  
Nerve Fibers ◽  
Ventral Horn ◽  
Double Labeling ◽  
Large Numbers ◽  
Simultaneous Visualization

Neurotransmitters have been shown to coexist in cell bodies, but demonstrating their coexistence within nerve fibers and terminals has been more difficult. However, two recent reports outlined a simple light-microscopic method by which two neurotransmitters can be shown to coexist in fibers and terminals. The method was identical to that used for immunohistochemical localization of one antigen, except that two primary--secondary antibody systems labeled with two different fluorochromes were used simultaneously. In the present study, a method for the simultaneous visualization of serotonin and substance P was characterized. This method employed an antiserum to serotonin generated in goat in combination with a rabbit-generated antiserum to substance P. These antisera were visualized with secondary antisera raised in swine and conjugated with rhodamine and fluorescein respectively. Spinal cord sections stained by this protocol showed large numbers of fibers fluorescing both red and green. Many of them were in the ventral horn, fewer were around the central canal, and virtually none were in the dorsal horn. The apparent double labeling could be shown not to be the result of cross-reactivity among the antisera, of any inappropriate affinity among the antisera, of green fluorescence by rhodamine, or of red fluorescence by fluorescein. It is concluded that the method provides a simple technique for visualizing fibers and terminals in which serotonin and substance P coexist.

Distribution and chemical coding patterns of cocaine- and amphetamine-regulated transcript-like immunoreactive (CART-LI) neurons in the enteric nervous system of the porcine stomach cardia

2015 ◽  
Vol 18 (3) ◽  
pp. 515-522 ◽  
Author(s):  
W. Rękawek ◽  
P. Sobiech ◽  
S. Gonkowski ◽  
K. Żarczyńska ◽  
A. Snarska ◽  
...  
Keyword(s):  
Nervous System ◽  
Enteric Nervous System ◽  
Neuronal Nitric Oxide Synthase ◽  
Nerve Fibers ◽  
Vesicular Acetylcholine Transporter ◽  
Double Labelling ◽  
Dense Network ◽  
Chemical Coding ◽  
Oxide Synthase ◽  
Labelling Immunofluorescence

Abstract The aim of this study was to determine the presence of cocaine- and amphetamine-regulated transcript-like immunoreactive (CART-LI) neurons and co-localisation of CART with vesicular acetylcholine transporter (VAChT), neuronal nitric oxide synthase (n-NOS), vasoactive intestinal polypeptide (VIP), substance P (SP) and leu-enkephalin (LENK) in the enteric nervous system of the porcine gastric cardia by using a double-labelling immunofluorescence technique. CART-LI neurons were observed in the myenteric plexus (18.2±2.6%). A dense network of CART-LI nerve fibers was mainly observed in the muscular layer. CART showed co-localization mainly with VAChT, n-NOS, VIP and to a lesser degree with LENK and SP. Distribution of CART and its co-localization with other neurotransmitters suggest that this peptide plays an important role in gastric motility in the pig.

scholarly journals Ultrastructural characterization of human large granular lymphocyte subsets defined by the expression of HNK-1 (Leu-7), Leu-11, or both HNK-1 and Leu-11 antigens.

1985 ◽  
Vol 33 (11) ◽  
pp. 1129-1133 ◽  
Author(s):  
G C Manara ◽  
G De Panfilis ◽  
C Ferrari
Keyword(s):  
Nk Cell ◽  
Double Labeling ◽  
Dense Granules ◽  
Ultrastructural Characterization ◽  
Nk Cell Differentiation ◽  
Round Nucleus ◽  
Immature Form ◽  
Irregular Cell ◽  
Leu 7

A peroxidase-colloidal gold double labeling system in immunoelectron microscopy was used to investigate the ultrastructural features of human large granular lymphocytes (LGL) subpopulations. Three subsets of LGL, Leu-7+-Leu-11-, Leu-7+-Leu-11+, Leu-7- -Leu-11+, were characterized using combinations of the monoclonal antibodies anti-Leu-7 and anti-Leu-11. They showed different ultrastructural patterns. In fact, Leu-7+-Leu-11- cells showed a high nuclear/cytoplasmic ratio (N/C), a round nucleus, a cytoplasm with few organelles, and a rather even surface. Moreover, most of them lacked electron-dense granules. On the other hand, Leu-11+ cells displayed a low N/C, an irregular-shaped nucleus, and a cytoplasm containing a well-developed Golgi apparatus, many mitochondria, vacuoles, vesicles, and numerous electron-dense granules. Moreover, they exhibited an irregular cell surface. Thus, Leu-7+-Leu-11- cells seemed to represent an immature form of LGL, while cells expressing the Leu-11 antigen showed a fine structure specific for functional NK cells. Our findings suggest that the expression of HNK-1 (Leu-7) and Leu-11 antigens respectively represents subsequent stages in NK cell differentiation.

scholarly journals The Influence of Bisphenol A (BPA) on Neuregulin 1-Like Immunoreactive Nerve Fibers in the Wall of Porcine Uterus

2018 ◽  
Vol 19 (10) ◽  
pp. 2962 ◽  
Author(s):  
Liliana Rytel
Keyword(s):  
Bisphenol A ◽  
Nerve Fibers ◽  
Double Immunofluorescence ◽  
Negative Effects ◽  
Protective Mechanisms ◽  
Neuregulin 1 ◽  
Small Doses ◽  
The Impact ◽  
First Time

Bisphenol A (BPA), a substance commonly used in the manufacture of plastics, shows multidirectional negative effects on humans and animals. Due to similarities to estrogens, BPA initially leads to disorders in the reproductive system. On the other hand, it is known that neuregulin 1 (NRG-1) is an active substance which enhances the survivability of cells, inhibits apoptosis, and protects tissues against damaging factors. Because the influence of BPA on the nervous system has also been described, the aim of the present study was to investigate for the first time the influence of various doses of BPA on neuregulin 1-like immunoreactive (NRG-1-LI) nerves located in the porcine uterus using the routine single- and double-immunofluorescence technique. The obtained results have shown that BPA increases the number and affects the neurochemical characterization of NRG-1-LI in the uterus, and changes are visible even under the impact of small doses of this toxin. The character of observed changes depended on the dose of BPA and the part of the uterus studied. These observations suggest that NRG-1 in nerves supplying the uterus may play roles in adaptive and protective mechanisms under the impact of BPA.

scholarly journals Newcastle disease virus may enter cells by caveolae-mediated endocytosis

2007 ◽  
Vol 88 (2) ◽  
pp. 559-569 ◽  
Author(s):  
Celia Cantín ◽  
Javier Holguera ◽  
Laura Ferreira ◽  
Enrique Villar ◽  
Isabel Muñoz-Barroso
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Inhibitory Effect ◽  
Endocytic Pathway ◽  
Host Cells ◽  
Virus Concentration ◽  
Double Labelling ◽  
Virus Adsorption ◽  
Labelling Immunofluorescence

The entry into cells of Newcastle disease virus (NDV), a prototype member of the paramyxoviruses, is believed to occur by direct fusion at the plasma membrane through a pH-independent mechanism. In addition, NDV may enter host cells by an endocytic pathway. Treatment of cells with drugs that block caveolae-dependent endocytosis reduced NDV fusion and infectivity, the degree of inhibition being dependent on virus concentration. The inhibitory effect was reduced greatly when drugs were added after virus adsorption. Cells treated with methyl β-cyclodextrin, a drug that sequesters cholesterol from membranes, reduced the extent of fusion, infectivity and virus–cell binding; this indicates that cholesterol plays a role in NDV entry. Double-labelling immunofluorescence assays performed with anti-NDV monoclonal antibodies and antibodies against the early endosome marker EEA1 revealed the localization of the virus in these intracellular structures. Using fluorescence microscopy, it was found that cell–cell fusion was enhanced at low pH. It is concluded that NDV may infect cells through a caveolae-dependent endocytic pathway, suggesting that this pathway could be an alternative route for virus entry into cells.

scholarly journals Cholinergic and VIPergic Innervation in Cerebral Arteries: A Sequential Double-Labeling Immunohistochemical Study

1990 ◽  
Vol 10 (1) ◽  
pp. 32-37 ◽  
Author(s):  
Frederick Jia-Pei Miao ◽  
Tony Jer-Fu Lee
Keyword(s):  
Vasoactive Intestinal Polypeptide ◽  
Arterial Wall ◽  
Immunohistochemical Study ◽  
Picric Acid ◽  
Cerebral Arteries ◽  
Nerve Fibers ◽  
Morphological Data ◽  
Immunohistochemical Method ◽  
Double Labeling ◽  
Functional Interactions

The possible co-localization of choline acetyltransferase (ChAT) and vasoactive intestinal polypeptide (VIP) in the nerve fibers of cat cerebral arteries was examined by a sequential double-labeling immunohistochemical method. Diaminobenzidine and tetramethylbenzidine were used as chromogens to distinguish ChAT (protein) and VIP (peptide) immunoreactivities. Since available fixatives often did not provide simultaneous preservation of optimal protein and peptide immunoreactivities, a new fixative, buffered periodate-paraformal-dehyde-picric acid-formaldehyde-lysine (PPPFL), was formulated and tested. PPPFL fixative is more reliable for simultaneously preserving ChAT and VIP immunoreactivities than were periodate-lysine-paraformaldehyde (PLP) fixative, Zamboni's fixative, or 2% paraformaldehyde solution alone. Using PPPFL as fixative, both ChAT immunoreactive (ChAT-I) and VIP-immunoreactive (VIP-I) fibers in cerebral arteries appeared as bundle and fine fibers. Most ChAT-I and VIP-I fibers were separate. Portions of ChAT-I and VIP-I fibers often ran closely in parallel or across each other. Overlaying of VIP-I on ChAT-I fibers and relay connections between them were also observed. These morphological data suggest the potential functional interactions between cholinergic and VIPergic innervations. In <5% of the fibers examined did ChAT and VIP immunoreactivities appear to be co-localized. These data therefore do not support the hypothesis that acetylcholine and VIP are co-localized in most fibers innervating the cerebral arterial wall.

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