Christo Chouchkov

Localization of somatostatin-like immunoreactive fibres in the trigeminal principal sensory nucleus of the cat

The location and distribution of nerve fibres and terminals displaying somatostatin-(SOM)-like immunoreactivity (LI) has been examined in cat trigeminal principal sensory nucleus (Vp) using the peroxidase-antiperoxidase technique. The varicose SOM-LI nerve fibres form a fine network in the Vp. SOM-positive terminals are distributed throughout Vp but they are particularly abundant in the ventral subdivision of the nucleus (Vpv). The labelled terminals make predominantly asymmetric axodendritic synaptic contacts. SOM-LI terminals only rarely form symmetrical synapses with non-reactive neuronal elements and are also present free in the neuropil. No SOM-LI perikarya are detected in the Vp. The results of the present study suggest that the majority, if not all, of SOM-LI fibres in the Vp are probably of primary afferent origin and may be involved in relaying trigeminal sensation to neuron located in this brain area.

Peptidergic innervation of the mesencephalic trigeminal nucleus in the cat.

The trigeminal processing of proprioceptive information is unique and very little is known about the neurochemical organization of trigeminal primary afferent neurons which mediate the sensory aspects of proprioception. In studies using immunocytochemical‐retrograde tracing techniques, some classical neurotramsitters mediating the afferent modulation of the mesencephalic trigeminal nucleus (MTN) have been investigated. This paper summarizes our current understanding of the peptidergic innervation of the cat MTN.

Ultrastuctural immunocytochemistry of calcium-binding proteins in rapidly-adapting avian mechanoreceptors

The localization of the calcium-binding proteins (CaBPs) calbindin-D28K (CB) and parvalbumin (PV) in avian rapidly-adapting Herbst and Grandry sensory corpuscles was studied with the use of immunocytochemistry and monoclonal antibodies. Strongest immunostaining was detected in cells of the capsule in both receptor types. Staining was more pronounced in the vicinity of endoplasmic reticulum and mitochondria in the perinuclear regions, whereas staining was distinct in pinocytotic vesicles in peripheral cytoplasmic lamellae. Fibroblasts and macrophages in the subcapsular space of Herbst receptors also showed strong immunostaining in organelles in perinuclear regions. Modified Schwann cells in both receptor types revealed moderately-expressed immunostaining, which was more pronounced in perinuclear regions. The various parts of the receptor nerve fibers showed weak to strong staining. The physiological roles of the investigated CaBPs may be associated with cytoplasmic calcium ion (Ca++) storage, which is necessary for either active metabolism in the immunostained structures and/or their transfer to sensory axonal regions where Ca++ channels are present.

Immunocytochemical localization of the neurons in the superior mesenteric ganglion innervating the small intestine of the cat

Retrograde tracing was used to determine the localization of neuronal perikarya and fibres in the feline superior mesenteric ganglion (SMG), projecting to the small intestine. In the distal part of the ileum, a retrograde neuronal tracer Fast Blue (FB) was injected and after approximately thirty five to forty days the animals were killed by perfusion. The SMG were removed and the neuropeptide contents of the neurons, projecting to the distal ileum, were determined by means of immunofluorescence with antisera to neuropeptide Y (NPY), calcitonin gene-related peptide (CGRP), substance P (SP), somatostatin (SOM), and vasoactive intestinal polypeptide (VIP). Neurons innervating the small intestine were located in the upper part of the SMG and all of them were NPY-immunopositive. The group of CGRP-immunoreactive (IR) cells was less numerous (73.33%). Probably the FB-labeled fibres, containing the same neuropeptides, arise from these perikarya. SP- or VIP-immunopositive neuronal processes were found to surround immunonegative ganglionic cells but their origin is not in the ganglion. Only single FB-marked cells were VIP-immunopositive. SP- and SOM-immunoreactive amounted respectively to 2.28% and 3.01% of all the neuronal population, but only a few of these cells were FB-labelled.

Localization of newly synthesized protein precursors of basement membrane in the embryonic central nervous system as revealed by radioautography

Using light- and electron microscope radioautography, the dynamics of newly synthesized protein precursors taking part in elaboration of brain basement membrane have been examined. The data presented give evidence that all cell types (endothelial, pericytal, and astroglial cells) surrounding the brain basement membrane contribute to its composition. A quantitative uptake analysis of 3H-proline indicates a progressive decline in the amount of labeled precursor in the examined cell types with a corresponding increase in deposition of the label at the presumptive basement membrane. The endothelial cells plays the main role in the membrane elaboration followed by pericyte and astrocyte which participate at lesser but equal degree in this synthesis.

Ruthenium red staining material in Herbst and Grandry sensory corpuscles.

The penetration and distribution of ruthenium red into Herbst and Grandry sensory corpuscles has been investigated. Ruthenium red positive substances have been localized in all intercellular spaces including the collagen fibrils, basement membranes and predominately the outer leaflet of the plasmalemmae including the axolemma. Some of the numerous pinocytotic and coated vesicles of the receptor cell layers have been heavily loaded with the dye. Sometimes, the intracytoplasmic and intraaxoplasmic penetration of ruthenium red has been observed. In these cases, the dye deposits have been restricted to the elements of smooth endoplasmic reticulum, mitochondria and microtubules.

Cytochemical localization of acid phosphatase and adenosine triphosphatase in some avian mechanoreceptors

Ultrastructural distribution of acid phosphatase and adenosine triphosphatase was studied in the receptor elements of HERBST and GRANDRY sensory corpuscles. Acid phosphatase activity was established in the elements of smooth and rough endoplasmic reticulum of perineural capsule cells, as well as in the secondary lysosomes of all cell types. Particular interest was paid on the activity of myelin-like dense bodies and some clear core vesicles belonging to the axoplasm of receptor nerve fibres. Adenosine triphosphatase activity was established on the membranes of receptor structures and pinocytotic vesicles. More deposits of electron dense material were localized on the axolemma of the non-myelinated portions of the receptor nerve fibres. The functional significance and importance of the both enzymes in the receptor structures was discussed.

Radioautographic study of leucine uptake and transmission of proteins by rat enterocytes following Concanavalin A binding.

The transport of labeled leucine and following protein synthesis has been investigated in rat enterocytes using quantitative electron microscope radioautography. The presented normal distribution of the label has been compared with the distribution of radioactivity after Concanavalin A treatment. It has been established that the transport of amino acid and subsequent newly synthesized proteins are delayed and confused after Concanavalin A binding.