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Experiments were designed to determine if neurons of the ranid optic tectum, a major target of the optic nerve, possess the same regenerative potential as optic axons. Normal tectal efferent (TE) projections were reexamined by using the anterograde transport of 3H-proline and autoradiography (n = 18), bulk-filling damaged TE axons with horseradish peroxidase (HRP; n = 18) and anterogradely transporting wheat germ agglutinin-HRP (n = 8) to label TE axons. Results were similar to reports that used degeneration methods (Rubinson: Brain Behav. Evol. 1:529-561, '68; Lazar: Acta. Biol. Hung. 20:171-183, '69). Following a brainstem hemisection just caudal to the nucleus isthmi (1-20 weeks), the ipsilateral descending TE pathway was autoradiographically examined (n = 20). While all other TE projections appeared normal, there was no detectable ipsilateral descending projection beyond the lesion site. Ascending TE axons were cut at the anterior tectal border by hemisecting the left diencephalon (LDH)--a lesion that also cuts optic axons projecting to the left tectum. There was no indication of TE axonal regeneration with the aid of autoradiography or HRP histochemistry 1-30 weeks postlesion (n = 48) even when the medial diencephalon was intentionally left intact (n = 4). However, in all four cases examined, optic axons regenerated following the same LDH where TE axonal regeneration failed (also see Stelzner, Lyon, and Strauss: Anat. Rec. 205:191A-192A, '83). Local effects of LDH should be similar for both the cut optic and cut TE axons. Other factors were tested that may contribute to the lack of TE axonal regeneration. Our results indicate that optic regeneration itself (n = 8), postaxotomy retrograde cell death of TE neurons (n = 6), deafferentation of the tectum of optic axons, and potential sprouting within tectal targets by intact contralateral TE axons (n = 10) are not critical factors aborting TE axonal regeneration. TE axons filled with HRP at chronic periods after LDH (n = 4) terminate anomalously near the LDH border. Many of these endings are similar to reactive endings or terminal clubs seen after axonal injury in the mammalian CNS. Our results suggest that this disparity in regenerative ability of optic and TE axons may be related to a difference in the responsive ability of these cell types to initiate or maintain axonal elongation after axotomy within the amphibian CNS environment.
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PMID:Tests of the regenerative capacity of tectal efferent axons in the frog, Rana pipiens. 302 86

During epididymal transit, the mouse sperm flagellum acquires a surface glycoprotein (SMA4) from epididymal fluid that functions as a sperm antiagglutinin. To determine the origin of this molecule, testes and epididymides of male mice were sectioned for light microscopy and stained with wheat germ agglutinin (WGA)-peroxidase, a probe that has been used previously to examine the biology of SMA4. WGA reactivity was localized to the cytoplasm in a small population of cells in the distal caput epididymis. Testis cells and principle cells of the caput were nonreactive with WGA, while stereocilia were stained on principle cells in the corpus and cauda. The WGA-positive cells in the distal caput were identified as holocrine cells on the basis of morphology, distribution, and PAS + reaction. At high magnification, intense WGA reactivity was due to the presence of numerous apical granules in the cytoplasm. The location of the cells in distal caput coincided exactly with the region of tubule in which sperm first acquired SMA4 on their flagellae. These data suggest that holocrine cells near the junction of caput and corpus epididymis are the source of the sperm antiagglutinin SMA4.
Anat Rec 1987 Feb
PMID:Maturation antigen of the mouse sperm flagellum: II. Origin from holocrine cells of the distal caput epididymis. 303 4

Density gradient centrifugation using a performed self generated gradient of colloidal silica enabled the isolation of microscopic sheep sarcocystis cystozoites, free from heart muscle contamination. The efficiency of separation of cystozoites from residual heart muscle after digestion in pepsin and hydrochloric acid was 63 to 92 per cent. Antigens from cystozoites were used on enzyme-linked immunosorbent assays (ELISA) of plasma from six coccidia-free lambs infected once orally with 70,000 microcystic sheep sarcocystis sporocysts and for raising antisera in rabbits. Use of an anti-sheep IgM conjugate in the ELISA showed that anti-sarcocystis IgM production was transitory, appearing five to 10 days after infection, peaking in concentration at 42 days and following the peak of the acute phase of infection (32 and 33 days) in the lambs. In contrast, total anti-sarcocystis immunoglobulins, detected by ELISA, increased from five to 21 days after infection and continued to increase until the lambs were killed (the last at 81 days) and was more useful in diagnosing chronic infection. No cross reactions between microcystic sheep sarcocystis and Toxoplasma gondii or Eimeria species of sheep were observed. A peroxidase anti-peroxidase test, using rabbit anti-sarcocystis sera, detected second generation meronts and sarcocysts in fixed tissues from infected lambs making it useful for the diagnosis of acute or chronic disease post mortem.
Vet Rec 1986 Nov 29
PMID:Experimental microcyst sarcocystis infection in lambs: serology and immunohistochemistry. 309 51

We have studied the response of nerve fibers containing calcitonin gene-related peptide immunoreactivity (CGRP-IR) to inflammation using a rate dental experimental system. Inflammation was induced by drilling tooth cusps to create pulpal exposures; the induced pulpitis and subsequent periapical lesions were studied 1-35 days later using standard CGRP immunohistochemistry and the avidin-biotin peroxidase method. The injury and resulting inflammation caused a disruption of CGRP-IR nerve fiber location and arborization that varied depending on whether the initial injury was limited to the pulp tip or extended throughout the pulp horn. At shorter survival periods (24 hr, 3 days) nerve fibers were either decreased or bundled into the center of the pulp with sprouting along the wound border. At 6 days necrosis and acute inflammation had advanced to varying degrees, and CGRP-IR fibers were extensively sprouted in the surviving pulp; the pulp also stained specifically for CGRP within 1-2 mm of the inflamed tissue at 6 days. At 35 days, we found total pulp necrosis in most teeth and the development of periapical bone loss, granulomatous tissue, and periapical abscesses. There was also an extensive increase in CGRP-IR nerve fibers in the tissues surrounding sites of severe periodontal inflammation and necrosis. In some cases, macrophage-like cells staining specifically for CGRP were near the abscesses. The results show important interactions between peptidergic nerve fibers and inflammatory cells, and are discussed in terms of the role of nerve fibers containing CGRP in neurogenic inflammation, mechanisms for intensification of CGRP immunoreactivity in affected fibers or neighboring cells, and implications for chronic inflammatory conditions, dental pain, and anesthesia.
Anat Rec 1988 Nov
PMID:Inflammation of rat molar pulp and periodontium causes increased calcitonin gene-related peptide and axonal sprouting. 326 42

The peroxidase anti-peroxidase technique was used to demonstrate free and intracellular immunoglobulin (Ig) within the lungs of 23 horses with chronic small airway disease. Histologically, all the horses had chronic bronchiolitis; however, the lesions varied in degree from mild in eight horses, to moderate in nine horses and severe in six horses. Lungs from three horses which had no gross or histopathological lesions were used as controls. In comparison with control horses, horses with mild chronic bronchiolitis had increased numbers of Ig A-containing and non-immunoglobulin staining cells around the vasculature and bronchioles. As the severity of the lesions increased so did the number of IgA-, IgG(Fc)- and in several cases non-immunoglobulin staining cells around the vasculature, bronchioles and in the alveolar septa. In severely affected horses, large amounts of free IgG(Fc) were observed interstitially and in alveoli. In areas of mucosal epithelial hyperplasia and metaplasia large amounts of free IgA and IgG(Fc) were sometimes observed interepithelially in a pattern which differed from that in control horses.
Vet Rec 1988 Feb 20
PMID:Chronic small airway disease in the horse: immunohistochemical evaluation of lungs with mild, moderate and severe lesions. 328 89

Expression of glycoconjugates during transfilter-induced differentiation of metanephric mesenchyme was studied by using fluorochrome- and peroxidase-coupled lectins. All cells in the uninduced metanephric mesenchyme expressed mannose, beta-D-galactose (beta-Gal), N-acetylglucosamine (GlucNAc), and terminal sialic acids. Additionally, solitary cells showed terminal alpha-D-galactose alpha-D-galactose (alpha-Gal) typical of mouse endothelial cells. During culture, undifferentiated mesenchymal cells seemed to disappear from induced explants, and many of the stromal cells between the evolving tubules presented terminal alpha-Gal residues. Similar positivity could be revealed in monolayer cultures of induced mesenchymes. A number of tubules in induced explants displayed alpha-L-fucosyl (Fuc) residues, characteristic of mature proximal tubules. Some terminal Ga1NAc residues, recognized only by Dolichos biflorus agglutinin, emerged in a few tubular cells after prolonged culture. The early tubules and glomerular bodies displayed a basement membrane presenting both terminal Ga1-(beta 1-3)-Ga1NAc and Ga1NAc residues. These positivities disappeared later from many tubular structures and glomerular bodies but persisted in tubules expressing proximal tubular differentiation. The glomerular bodies displayed only one cell type, reminiscent of maturing podocytes, presenting terminal Ga1-(beta 1-3)-Ga1NAc and Ga1NAc residues. Later these saccharide residues became covered by sialylation, as they could then be revealed only after treatment with neuraminidase. The results indicate that the segment-specific expression of saccharide residues during differentiation of nephron in vitro resembles the sequence seen in vivo. This study also suggests that the basement membranes surrounding the nephron show a stepwise, segment-specific maturation. Despite the presence of endothelial cells in the metanephric explants, only avascular glomeruli evolved in this differentiation model.
Anat Rec 1988 Feb
PMID:Expression of cellular glycoconjugates in transfilter-induced metanephric mesenchyme. 335 61

In the small intestine, the presence of transitional cells or cells intermediate between Paneth cells and goblet cells has been reported frequently for 100 years. Light microscopy and, more recently, fine structural studies have indicated that secretory granules observed in intermediate cells share some morphologic characteristics with those of granular goblet cells and of Paneth cells. In order to verify if intermediate cells in the jejunum and ileum of the adult mouse have functional similarities with either granular goblet or Paneth cells, we have studied the incorporation of sulfur-35 by radioautography and the localization of lysozyme by immunocytochemistry. After radioautography, goblet cells and, to a lesser extent, granular goblet cells had incorporated sulfur-35, whereas Paneth cells and intermediate cells were completely negative. Immunolocalization of lysozyme was done by using rabbit anti-rat lysozyme and protein A-peroxidase. After demonstration of peroxidase activity, only Paneth cells were stained and intermediate cells were negative. Therefore, intermediate cells do not contain sulfomucin or lysozyme, and they are functionally different from goblet and Paneth cells. Their function remains unknown.
Anat Rec 1988 Mar
PMID:On the presence of intermediate cells in the small intestine. 336 55

Ultrastructural, functional, and cytochemical characteristics of resident sinusoidal macrophages (RSM) in brown bullhead (Ictalurus nebulosus) liver were examined. Following perfusion fixation of the hepatic vascular bed, light micrographs revealed RSM that possessed multiple elongate cytoplasmic processes and frequently contained erythrocytes in various stages of degradation. Following brief perfusion fixation, light microscope examination of vibratome sections of bullhead liver reacted for peroxidase revealed intensely positive RSM. By transmission electron microscopy, peroxidase activity was localized to the nuclear envelope and cytoplasmic granules of RSM and in endothelial and perisinusoidal fat-storing cells. In cryostat sections of fresh-frozen liver, glucose-6-phosphate dehydrogenase (G-6-PDH) was uniformly distributed over hepatocytes, whereas intensely positive punctate staining for G-6-PDH was localized over RSM. To test for phagocytosis by RSM, latex beads (0.81 micron) were injected into a tributary of the hepatic portal vein 2 min prior to perfusion fixation. Latex beads appeared either singly or in dense aggregates within RSM. Ultrastructurally, RSM were characterized by an irregularly shaped, eccentrically located nucleus, electron-dense vacuoles, small patches of granular endoplasmic reticulum, a well-developed Golgi apparatus, elongated mitochondria, desmosomes or desmosome-like densities that served as a source of attachment to endothelial cells, and a centriole with radiating microtubules. Invaginations of the plasma membrane (vermiform processes) characteristic of mammalian Kupffer cells were not observed in bullhead RSM. The results indicated a resident cell population of sinusoidal macrophages in the bullhead liver with properties that partially resembled mammalian Kupffer cells. These results are important for the identification of the normal resident cells in the bullhead liver.
Anat Rec 1987 Dec
PMID:Resident sinusoidal macrophages in the liver of the brown bullhead (Ictalurus nebulosus): an ultrastructural, functional and cytochemical study. 344 51

Although the reinnervation of muscle grafts has been demonstrated, the question remains as to the source of neurons reinnervating the graft. A muscle graft could be reinnervated by its original neurons (neural neurotization) or by sprouting of axons which supply the intact myofibers of surrounding muscles (muscle neurotization). The objective of this study was to evaluate the role of muscle neurotization in the reinnervation process using both horseradish peroxidase and fluorescent tracers. Observations from 20 muscle grafts indicate that the neurons reinnervating the grafts are from the original motor neuron pool. Thus muscle neurotization may not occur during the reinnervation process.
Anat Rec 1987 Dec
PMID:Role of muscle neurotization in the reinnervation of murine muscle grafts. 344 53

A protein of approximately 28,000 relative molecular mass (Mr) cross-reacting with antiserum against the 28,000-Mr rat renal calcium-binding protein (calbindin-D28k) has been localized in the kidney of a salientian amphibian, Rana catesbeiana. Cells reactive for calbindin-D28k were found in the distal tubule at all stages of metamorphosis by the unlabeled antibody peroxidase-antiperoxidase technique. Adult kidneys appeared to have more calbindin-D28k-positive cells. The renal corpuscle, neck, and proximal tubule were negative. An immunoreactive 28,000-Mr band that comigrated with the band of calbindin-D28k was visualized by the immunoblot technique. The finding of the 28,000-Mr calbindin-D in the anamniotic kidney demonstrates that this calcium-binding protein (CaBP) is phylogenetically older than our previous studies of higher vertebrates had revealed (Rhoten et al., 1985). Although the function of calbindin-D28k in the distal nephron is unknown, this CaBP can now be presumed to have functional significance in the mesonephric as well as the metanephric kidney.
Anat Rec 1986 Oct
PMID:Calcium-binding protein (28,000 Mr calbindin-D28k) in kidneys of the bullfrog Rana catesbeiana during metamorphosis. 349 Aug 5


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