Immunohistochemical characterization of neurons in the vestibular ganglion (Scarpa’s ganglion) of the pig

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

Morphology and immunohistochemical characteristics of the pterygopalatine ganglion in the chinchilla (Chinchilla laniger, Molina)

Histological and histochemical investigations revealed that the pterygopalatine ganglion (PPG) in the chinchilla is a structure closely connected with the maxillary nerve. Macro-morphological observations disclosed two different forms of the ganglion: an elongated stripe representing single agglomeration of nerve cells, and a ganglionated plexus comprising smaller aggregations of neurocytes connected with nerve fibres. Immunohistochemistry revealed that nearly 80% of neuronal cell bodies in PPG stained for acetylcholine transferase (CHAT) but only about 50% contained immunoreactivity to vesicular acetylcholine transporter (VACHT). Many neurons (40%) were vasoactive intestinal polypeptide (VIP)-positive. Double-staining demonstrated that approximately 20% of the VIP-immunoreactive neurons were VACHT-negative. Some neurons (10%) in PPG were simultaneously VACHT/nitric oxide synthase (NOS)- or Met-enkephaline (Met-ENK)/CHAT-positive, respectively. A small number of the perikarya stained for somatostatin (SOM) and solitary nerve cell bodies expressed Leu-ENK- and galanin-immunoreactivity. Interestingly about 5-8% of PPG neurons exhibited immunoreactivity to tyrosine hydroxylase (TH). Intraganglionic nerve fibres containing immunoreactivity to VACHT-, VIP- and Met-ENK- were numerous, those stained for calcitonin gene related peptide (CGRP)- and substance P (SP)- were scarce, and single nerve terminals were TH-, GAL-, VIP- and NOS-positive

Immunohistochemical properties of motoneurons supplying the trapezius muscle in the rat

Combined retrograde tracing (using fluorescent tracer Fast blue) and double-labelling immunofluorescence were used to study the distribution and immunohistochemical characteristics of neurons projecting to the trapezius muscle in mature male rats (n=9). As revealed by retrograde tracing, Fast blue-positive (FB+) neurons were located within the ambiguous nucleus and accessory nucleus of the grey matter of the spinal cord. Immunohistochemistry revealed that nearly all the neurons were cholinergic in nature [choline acetyltransferase (ChAT)-positive]. Retrogradely labelled neurons displayed also immunoreactivities to calcitonin gene-related peptide (CGRP; approximately 60% of FB+ neurons), nitric oxide synthase (NOS; 50%), substance P (SP; 35%), Leu5-Enkephalin (LEnk; 10%) and vasoactive intestinal polypeptide (VIP; 5%). The analysis of double-stained tissue sections revealed that all CGRP-, VIP- and LEnk-immunoreactive FB+ perikarya were simultaneously ChAT-positive. The vast majority of the neurons expressing SP- or NOS-immunoreactivity were also cholinergic in nature; however, solitary somata were ChAT-negative. FB+ perikarya were surrounded by numerous varicose nerve fibres (often forming basket-like structures) immunoreactive to LEnk or SP. They were also associated with some CGRP-, NOS- and neuropeptide Y-positive nerve terminals

Caudal mesenteric ganglion in the sheep-macroanatomical and immunohistochemical study

The caudal mesenteric ganglion (CaMG) is a prevetrebral ganglion which provides innervation to a number of organs in the abdominal and pelvic cavity. The morphology of CaMG and the chemical coding of neurones in this ganglion have been described in humans and many animal species, but data on this topic in the sheep are entirely lacking. This prompted us to undertake a study to determine the localization and morphology of sheep CaMG as well as immunohistochemical properties of its neurons. The study was carried out on 8 adult sheep, weighing from 40 to 60 kg each. The sheep were deeply anaesthetised and transcardially perfused with 4% paraformaldehyde. CaMG-s were exposed and their location was determined. Macroanatomical observations have revealed that the ovine CaMG is located at the level of last two lumbar (L5 or L6) and the first sacral (S1) vertebrae. The ganglion represents an unpaired structure composed of several, sequentially arranged aggregates of neurons. Immunohistochemical investigations revealed that nearly all (99.5%) the neurons were DßH-IR and were richly supplied by VACHT-IR nerve terminals forming „basket-like" structures around the perikarya. VACHT-IR neurones were not determined. Many neurons (55%) contained immunoreactivity to NPY, some of them (10%) stained for Met-ENK and solitary nerve cells were GAL-positive. CGRP-IR nerve fibres were numerous and a large number of them simultaneously expressed immunoreactivity to SP. Single, weakly stained neurones were SP-IR and only very few nerve cells weakly stained for VIP

Zebrafish: an animal model for research in veterinary medicine

The zebrafish (Danio rerio) has become known as an excellent model organism for studies of vertebrate biology, vertebrate genetics, embryonal development, diseases and drug screening. Nevertheless, there is still lack of detailed reports about usage of the zebrafish as a model in veterinary medicine. Comparing to other vertebrates, they can lay hundreds of eggs at weekly intervals, externally fertilized zebrafish embryos are accessible to observation and manipulation at all stages of their development, which makes possible to simplify the research techniques such as fate mapping, fluorescent tracer time-lapse lineage analysis and single cell transplantation. Although zebrafish are only 2.5 cm long, they are easy to maintain. Intraperitoneal and intracerebroventricular injections, blood sampling and measurement of food intake are possible to be carry out in adult zebrafish. Danio rerio is a useful animal model for neurobiology, developmental biology, drug research, virology, microbiology and genetics. A lot of diseases, for which the zebrafish is a perfect model organism, affect aquatic animals. For a part of them, like those caused by Mycobacterium marinum or Pseudoloma neutrophila, Danio rerio is a natural host, but the zebrafish is also susceptible to the most of fish diseases including Itch, Spring viraemia of carp and Infectious spleen and kidney necrosis. The zebrafish is commonly used in research of bacterial virulence. The zebrafish embryo allows for rapid, non-invasive and real time analysis of bacterial infections in a vertebrate host. Plenty of common pathogens can be examined using zebrafish model: Streptococcus iniae, Vibrio anguillarum or Listeria monocytogenes. The steps are taken to use the zebrafish also in fungal research, especially that dealing with Candida albicans and Cryptococcus neoformans. Although, the zebrafish is used commonly as an animal model to study diseases caused by external agents, it is also useful in studies of metabolic disorders including fatty liver disease and diabetes. The zebrafish is also a valuable tool as a model in behavioral studies connected with feeding, predator evasion, habituation and memory or lateralized control of behavior. The aim of the present article is to familiarize the reader with the possibilities of Danio rerio as an experimental model for veterinary medicine