Immunohistochemistry of biogenic polypeptides in nerve cells and fibres of the guinea pig inferior mesenteric ganglion after perturbations

Summary Immunohistochemistry of peptide-and dopamine--hydroxylase-(DBH)-containing varicose nerve fibres and ganglion cells, respectively, in the guinea pig inferior mesenteric ganglion was investigated followinga) transsection of mesenteric (colonic) branches,b) transsection of central (lumbar splanchnic, intermesenteric and hypogastric) branches, andc) transplantation into the spleen.The findings indicate that pathways of different opioid peptides are not identical Met-enkephalin-and met-enkephalin-arg-phe-(cleavage products from pre-proenkephalin) containing fibres course in central branches to make contact in the inferior mesenteric ganglion. Dynorphin-and -neo-endorphin-(deriving from pre-prodynorphin) containing fibers as well as leu-enkephalin-(included in the dynorphin sequence) fibres appear to rise not only from central and from enteric somata, but also from intraganglionic noradrenergic neurons. Similar pathways seem to be used by VIP-and by neurotensin-immunoreactive cell bodies are rare. Practically all substance P-and most CGRP-immunoreactive fibres enter the ganglion via central branches and, to a large extent, traverse it, but some CGRP-immunoreactive influx appears to come from the intestine. The origin of intraganglionic substance P-and CGRP-immunoreactive fibres after ganglion transplantation remained unidentified. Somatostatin-and neuropeptide Y-immunoreactive fibres predominantly have an intraganglionic origin as have DBH-immunoreactive noradrenergic fibres. The demonstrated alterations in neuropeptide immunoreactivity of intraganglionic and periganglionic nerve fibres following the applied transsection procedures contribute to the present knowledge on origin and destination of peptidergic transmitter segments in the guinea pig inferior mesenteric ganglion. Moreover, the present study provides evidence that intrinsic participitation in intraganglionic fibre supply is more extensive than hitherto believed.Dedicated to Professor Dr. T.H. Schiebler on the occasion of his 65th birthdaySupported by the Deutsche Forschungsgemeinschaft, grant He 919/6-2     

Immunohistochemistry of biogenic polypeptides in nerve cells and fibres of the guinea pig inferior mesenteric ganglion after perturbations

Immunohistochemistry of peptide- and dopamine-beta-hydroxylase-(DBH)-containing varicose nerve fibres and ganglion cells, respectively, in the guinea pig inferior mesenteric ganglion was investigated following a) transsection of mesenteric (colonic) branches, b) transsection of central (lumbar splanchnic, intermesenteric and hypogastric) branches, and c) transplantation into the spleen. The findings indicate that pathways of different opioid peptides are not identical. Met-enkephalin- and met-enkephalin-arg-phe- (cleavage products from pre-proenkephalin) containing fibres course in central branches to make contact in the inferior mesenteric ganglion. Dynorphin- and alpha-neo-endorphin- (deriving from pre-prodynorphin) containing fibres as well as leu-enkephalin- (included in the dynorphin sequence) fibres appear to rise not only from central and from enteric somata, but also from intraganglionic noradrenergic neurons. Similar pathways seem to be used by VIP- and by neurotensin-immunoreactive fibres, although intraganglionic neurotensin-immunoreactive cell bodies are rare. Practically all substance P- and most CGRP-immunoreactive fibres enter the ganglion via central branches and, to a large extent, traverse it, but some CGRP-immunoreactive influx appears to come from the intestine. The origin of intraganglionic substance P- and CGRP-immunoreactive fibres after ganglion transplantation remained unidentified. Somatostatin- and neuropeptide Y-immunoreactive fibres predominantly have an intraganglionic origin as have DBH-immunoreactive noradrenergic fibres. The demonstrated alterations in neuropeptide immunoreactivity of intraganglionic and periganglionic nerve fibres following the applied transsection procedures contribute to the present knowledge on origin and destination of peptidergic transmitter segments in the guinea pig inferior mesenteric ganglion. Moreover, the present study provides evidence that intrinsic participation in intraganglionic fibre supply is more extensive than hitherto believed.     

Spermiogenesis in Lumbricus herculeus; an electron microscope study

Small pieces of the sperm sacs of Lumbricus herculeus were fixed for 4 hours in chrome-osmium, embedded in methacrylate, sectioned with a Porter-Blum microtome, and studied with a R.C.A. EMU-2C electron microscope. Each spermatid of a group developing synchronously is attached by a cytoplasmic strand to a common nutrient protoplasmic mass. This mass contains mitochondria and yolk bodies but is anucleate. The proximal centriole, that is, the centriole nearer the nucleus, is at first associated with a small peg which becomes firmly attached to the nuclear membrane. Later these two bodies become separated during the development of the middle-piece which is differentiated in the usual manner from a nebenkern formed by the fusion of 6 or 7 mitochondria. The acrosome develops in relation to the dictyosome (Golgi body), itself composed of 8 or more individual flattened sacs and situated in the cytoplasm opposite the point of attachment of the spermatid to the nutrient mass. Soon after its formation, the acrosome becomes incorporated into a cytoplasmic appendage or acrosome carrier. The carrier moves from its original position, along the lateral border of the elongating nucleus, to the distal margin of the nucleus where the acrosome is deposited. No evidence was found of a centriole located at the point of junction between nucleus and acrosome as suggested by earlier workers.     

An electron microscope study of the regenerating nerve fibers

Summary Guinea pig sciatic nerves were severed in order to obtain a regenerative process of the nerve fibers. The animals were killed at different periods of time after the severing (24 hours, 6 days and 12 days) and the specimens obtained were prepared for electronmicroscopic study.The nerve fiber growing extremities (growing cones) were specially studied. The growing cones showed the following components: a) microvesicles; b) mitochondria; c) multivesicular bodies.The microvesicles are hollow elements of about 200 to 700 Å. They constitute the main component of the growing cone. The mitochondria were seen as elongated bodies of 80 m. They were seen in many cases changing to round dense bodies which appear to break-up in irregularly-shaped fragments.The multivesicular bodies were found present in most of the growing cones. Protoneurofibrils do not exist in the growing cone but a close relationship between microvesicles and protoneurofibrils was found in the segments next to the growing cone.The above-mentioned components were found in all growing cones, disregarding the time elapsed after the nerve severing.Director of the Institute.Assistant of the Department of Neurohistology and Experimental Histology.Head of the Department of Cell Ultrastructure.     

Localization of substance P and leucine enkephalin in the nerve terminals of the guinea pig paracervical ganglion

Summary Immunoreactivities (IR) of substance P and leucine enkephalin have been demonstrated in the guinea-pig paracervical ganglion by an immunogold electron microscope method. Both substance P-IR and leucine enkephalin-IR were detected in large synaptic vesicles with electron-dense cores. The former neuropeptide was detected in nerve terminals and varicosities comprised mainly of large vesicles with electron-dense cores; the latter was detected in nerve terminals and varicosities that also included small, clear synaptic vesicles. In a minority of nerve terminals and varicosities coexistence of both immunoreactivities could be demonstrated within vesicles with an electron-dense core. Also present in these nerve terminals and varicosities were small, clear synaptic vesicles, though these were unreactive.     

Light microscopy study of Schmidt-Lantermann incisures in the guinea pig sciatic nerve

The authors studied the shape, orientation and number of Schmidt-Lantermann incisures in the sciatic nerve of the healthy guinea pig. The nerves were removed, fixed, shredded and evaluated in the light microscope. Internode length, fibre diameter and segment length were measured in isolated nerve fibres with an eyepiece micrometer. The number and configuration of the Schmidt-Lantermann incisures in the internode were found to be variable. The incisures follow one another at irregular intervals, dividing the whole internode into cylindroconical segments of varying lengths. The measured values were processed on a PMD 85/2 personal microcomputer.     

Histopathological changes in the eyes in systemic lupus erythematosus: an electron microscope and immunohistochemical study

This paper reports the histopathological findings in the eyes of a 26-year-old female patient diagnosed with systemic lupus erythematosus (SLE) with peripheral neuropathy. The patient had no significant ocular problems. She died of pneumonia after two years of suffering. The eyeballs were procured at autopsy and the retina, choroid and optic nerve processed for light and electron microscopy, and immunohistochemistry for immunoglobulin G (IgG), glial fibrillary acidic protein (GFAP), calbindin and parvalbumin. Histologically, there was haemorrhaging in the retinal nerve fibre layer. Ultrastructurally, the axons of this layer were swollen, and contained an unusual accumulation of microtubules and smooth endoplasmic reticulum. There were degenerative changes in the pericytes and smooth muscle cells of blood vessels. The capillary lumen was partially obliterated, and contained IgG, which was also detected throughout the choroid and wall of choroidal arterioles. The latter and Bruch's membrane showed fibrin deposits. The optic nerve showed infiltrated mononuclear cells near the degenerated axons, these axons lacked immunoreactivity to calbindin and parvalbumin. Compared to the control, the connective tissue sheaths of the central retinal vessels possessed a vast number of proliferated fibroblast cells, and trichrome staining showed transmural vessel scarring. Dense GFAP immunoreactivity was observed surrounding the vessel wall. These pathological changes are due to impaired blood circulation caused by haemorrhaging and vasculitis, and vessel occlusion by fibrin. The nature of the changes observed tends to indicate that a regular, thorough ophthalmic examination should be conducted even in the absence of significant ocular symptoms in SLE.     

Electron microscope studies of experimentalEntamoeba histolytica infection in the guinea pig

Early cellular and vascular changes in response to invasion of lamina propria byEntamoeba histolytica were studied sequentially, at the ultrastructural level, in germfree guinea pigs inoculated intracecally with amebae and enteric flora derived from patients with acute amebic colitis. Approximately one week post-inoculation the animals developed acute colitis with mucosal invasion by trophic amebae. Although epithelial cells at the sites of amebic invasion showed progressive cytoplasmic changes and desquamation resulting in microerosions, most mesenchymal elements in the lamina propria appeared normal without cytopathic changes even when in direct contact with invading amebae. Only the polymorpho-nuclear leukocytes (PMN) apposed or topographically close to amebae exhibited degenerative changes which were characterized by condensation of nucleoplasm and cytoplasm, extracellular release of cytoplasmic components including granules, and, finally, lysis of cell membranes. Capillaries and venules in the lamina propria showed a variety of changes such as swelling and gap formation at the intercellular endothelial junctions and more rarely at the fenestrae. Blood vessels physically close to amebae showed formation of endothelial cytoplasmic blebs which pinched off into the vascular or extravascular space. Platelet and fibrin thromboses were common in the more severely damaged capillaries and venules. Fragments or clumps of fibrin-like material were found also in the extracellular spaces. Amebic invasion of the lamina propria, then, is accompanied by continued epithelial shedding, PMN degeneration, and changes in both capillaries and venules consisting of endothelial damage and occlusive thrombosis. The vascular changes appeared to be closely related to PMN degeneration resulting from interaction of PMN with invading amebae.     

Electron microscope study of the developing nerve terminals in the acoustic organs of the chick embryo

Summary The stages of growth of the acoustic pathway (peripheral branch) were studied with the electron microscope in serial sections of the acoustic organs of 3 to 7 day chick embryos.Migration of cells from the acoustic epithelium was found at three days of incubation. These cells are presumably the futur ganglion cells. Fascicles of nerve fibers penetrate the epithelium through gaps of the basement membrane at 4–5 days of incubation. A dilatation develops in the intraepithelial fibers at about six days and thin and long prolongations grow from these dilatations and distribute among the cells. In the course of the next day the fibers embrace the foot of the sensory cell and the prolongations become shortened. Many of these extensions are charged with vesicles. At this stage (seven days) specialized structures (synaptic bars) differentiate in the region of the sensory cell contacting the large nerve ending (calix) or its short extensions. Each cell may show several synaptic bars, and each prolongation may contact with more than one cell.Research sponsored by the Air Force Office of Scientific Research, Office of Aerospace Research, United States Air Force, under AFOSR Grant Nr. 313-67.     

Structural changes of the guinea pig prostatic epithelial cells after gossypol treatment

The effect of gossypol acetic acid on the prostate gland of the guinea pig was assessed ultrastructurally. The three lobes of the gland showed differences in sensitivity and reacted differently to gossypol treatment. In the lateral prostate, there was a reduction in the profile of the granular endoplasmic reticulum and dense secretory granules with a concurrent appearance of smaller clear granules and an increase in cytoplasmic filamentous bundles. The latter feature was similar to that of the coagulating gland. In the dorsal prostate, the general reaction was similar to that in the lateral prostate except that there was no increase in filamentous content. In the coagulating gland, there was a reduction or total disappearance of apical secretory blebs and an increase in lysosomes, a feature not found in the lateral and dorsal lobes. Damages to mitochondria were observed in the dorsal prostate and coagulating gland but not in the lateral prostate. It is concluded that gossypol affects not only the testis, but also alters the structure and functions of the prostate gland.     

Innovar-Vet-induced pathologic changes in the guinea pig.

High doses of Innovar-Vet administered im induced pathologic changes in tissues at the site of drug deposition. The onset and extent of the changes were dose related. Distal self-mutilation occurred coincidentally to the peak of pathologic changes in the ischiatic nerve.