Guinea Pig Endolymphatic Sac Research Articles

Electrocochleography in Experimental Endolymphatic Hydrops

This study investigated whether dominant negative summating potential (DNSP) is absent at all stages of induced hydrops development, including the early stages of hydrops formations. Electrocochleography (ECoG) was done 3 days to 20 weeks after obliteration of the endolymphatic sac in guinea pigs, by electrodes attached to the cochlear bony wall on the scala vestibuli of the basal turn. DNSP was noticed only during the early stages of endolymphatic hydrops formation, before hydrops was fully developed. There was no DNSP when the distension of Reissner's membrane was marked. Increased endolymphatic pressure and/or changes in biochemical composition were thought to be the causes of DNSP.

S-phase cells in the guinea pig endolymphatic sac.

S-phase (the period of DNA synthesis) cells in the endolymphatic sac (ES) were investigated by immunohistochemical methods. Monoclonal antibodies against 5-bromo-2'-deoxyuridine (BrdUrd) and proliferating cell nuclear antigen (PCNA) were used. After administration of BrdUrd i.p., guinea pigs were sacrificed with acid formalin and the decalcified temporal bones examined immunohistochemically. Scattered cells labelled with anti-BrdUrd or anti-PCNA antibodies were detected within the ES whereas labelled cells were not found in either the cochlea or the vestibule. From the morphological findings, these labelled cells within the ES epithelium were considered to be epithelial cells and the sub-epithelial stained cells looked like fibroblastic cells. Some labelled resident floating cells were found in the ES lumen. Our results reveal that the ES, where resident cells proliferate, is the sole site of the cell-renewing system in the inner ear of an adult animal.

Na,K-ATPase subunit isoform expression in the guinea pig endolymphatic sac.

Na,K-ATPase subunit isoform expression was studied by immunocytochemistry in the guinea pig endolymphatic sac, using subunit isoform-specific polyclonal antibodies. Epithelial cells of the guinea pig endolymphatic sac were observed to contain the alpha 1- and beta 2-subunit isoforms, and to a lesser extent the beta 1-subunit isoform, of Na,K-ATPase. The alpha 1- and beta 2-subunit isoforms of Na,K-ATPase have been observed previously in other ion and fluid transporting regions of the membranous labyrinth, e.g., stria vascularis and vestibular dark cells. Combined data indicate that the alpha 1-, beta 2-form of Na,K-ATPase plays a role in the microhomeostasis of endolymph. The alpha 1 beta 2 Na,K-ATPase subunit isoform combination is different from typical ion and fluid transporting tissues, e.g., kidney and colon, and may reflect distinctive characteristics of inner ear Na,K-ATPase.

Sympathetic and CGRP-positive nerve supply to the endolymphatic sac of guinea pig.

The distribution of sympathetic fibers and calcitonin gene related peptide (CGRP)-positive fibers was examined in the endolymphatic sac of 10 guinea pigs by using immunohistochemical techniques to demonstrate tyrosine hydroxylase (TH) and CGRP. A meshwork of TH-positive fibers, found around the sigmoid sinus, sent branches to the distal and the intermediate parts of the endolymphatic sac. In these areas. TH-positive fibers traveled freely from blood vessels and formed a loose plexus beneath the lining epithelium. Such fibers were rare in proximal portion of the endolymphatic sac and in the endolymphatic duct. Seven days after elimination of the ipsilateral superior cervical ganglion, some of the TH-positive fibers were gone, however, there were still a few fibers in the endolymphatic sac as well as around the sigmoid sinus, thus their origin remains obscure. CGRP-positive fibers also branched from the fibers around the sigmoid sinus, and were distributed throughout the endolymphatic sac, some occasionally extending to the endolymphatic duct. They not only formed a dense plexus in the sublining space, but also spread through the lining cell layer. Elimination of the ipsilateral superior cervical ganglion did not affect the distribution of CGRP-positive fibers, indicating that they are probably not sympathetic fibers.

Immunohistochemical localization of carbonic anhydrase in the endolymphatic sac of the mouse and guinea pig.

The localization of carbonic anhydrase (CA) in mouse and guinea pig endolymphatic sac (ES) was determined by immunohistochemistry. In the distal and intermediate portions of the ES of both species, epithelial cells reacted with anti-CA antiserum. All epithelial cells in the distal portion stained for CA. In the intermediate portion, immunoreactivity was prominent in about half the epithelial cells, but less intense in the remaining epithelial cells. The proximal portion of the ES lacked immunoreactivity. CA may play a crucial role in ES electrolyte transport.

Hyaluronan synthesis in the adult guinea pig endolymphatic sac.

The endolymphatic sac is believed to play a major role in membranous labyrinth homeostasis by controlling the volume of endolymph, removing debris, and participating in the immune response of the inner ear. The endolymphatic sac is postulated to absorb endolymph and to synthesize and secrete high-molecular-weight and osmotically active glycosaminoglycans (GAGs). The present study examines the ability of in vitro adult guinea pig endolymphatic sac cells to synthesize complex proteins and polysaccharides. The intent is to characterize the nature of these compounds by studying carbon-14 (14C) glucose incorporation in tissue cultured endolymphatic sac specimens using autoradiographic and specific enzymatic digestion techniques. Our results suggest that sac cells can synthesize GAGs and proteins in vitro in proportionately larger amounts than surrounding connective tissue and dura. The principal GAG synthesized by the endolymphatic sac appears to be hyaluronan.

Development of endolymphatic hydrops following immune response in the endolymphatic sac of the guinea pig.

The present study investigated immune injury associated with endolymphatic hydrops (e.hydrops) following locally mounted immune reaction in the endolymphatic sac (e.sac) of guinea pigs. E.hydrops occurred, progressing rapidly within the first week post secondary Keyhole limpet hemocyanin (KLH) challenge in the e.sac and developed into two phases, acute and chronic. On the other hand, primary KLH challenge of the e.sac, PBS inoculation into the e.sac or intradural secondary KLH challenge were incapable of inducing e.hydrops. These results indicate that reversible and irreversible e.hydrops are induced by the immune response of the e.sac, suggesting that local immunological events of the e.sac may provide an animal model of Meniere's disease.

Electrocochleography in Guinea Pigs with Experimental Endolymphatic Hydrops

In this study we attempt to determine whether the dominant negative summating potential (DNSP) is present during the earlier stages of induced hydrops development. Electrocohleographs (ECoG) were taken 3 days, 4 days, 1 week, 2 weeks, 3 weeks, and 10-20 weeks after obliteration of the endolymphatic sac in guinea pigs with electrodes in the basal turn scala vestibuli.The action potential (AP) thresholds rose and the AP amplitudes decreased with time following surgery. The increases in the summative potential (SP) amplitudes and in the rises in the SP/AP ratios at 95 dB pe SPL in response to clicks tended to be greater 3 and 4 days postoperatively than subsequently. These results indicate that the potential for DNSP exists during the early stages of endolymphatic hydrops formation, before the hydrops is fully developed.

Calmodulin binding sites in the endolymphatic sac and stria vascularis of the human fetus and the guinea pig.

Calmodulin (CaM) binding sites, which are available and unoccupied by CaM at the time of fixation, were analyzed in the guinea pig endolymphatic sac and stria vascularis using CaM conjugated tetramethyl rhodamine isothiocyanate. The endolymphatic sac epithelium and stria vascularis both showed strong fluorescence. Weak CaM binding sites were seen in the endolymphatic sac in the 12-week-old human fetus. In contrast, the fluorescent reaction in the stria vascularis and endolymphatic sac in 14- and 15-week-old human fetuses was strong. These results suggest that CaM may play an important role for the maintenance of ionic stability in the endolymph.

Effect of Ototoxic Drug Administration to the Endolymphatic SAC

Aminoglycosides were applied to the endolymphatic sac of guinea pigs, and the inner ears were studied by light microscopy. In group 1 gentamicin sulfate application on the lateral surface of the endolymphatic sac produced sensory cell atrophy limited to the macula sacculi. In group 2 gentamicin injection into the sac by extradural and intradural approaches often showed lesions in the maculae sacculi and to the sac from a lateral opening resulted in a similar pattern of degeneration but with less severity in comparison to the injection method. Mild to severe endolymphatic hydrops was sometimes observed. The study shows diffusion of drugs taking place against the longitudinal flow of endolymph. This procedure may be applicable for the treatment of Meniere's disease.

内リンパ嚢局所免疫モルモットの前庭機能障害について

We examined vestibular dysfunction and histological damage caused by direct antigen challenge to the endolymphatic sac in guinea pigs. We observed spontaneous nystagmus every eight hours and performed caloric testing every week following endolymphatic sac secondary KLH challenge. Spontaneous nystagmus was seen in 12 of 18 animals, and nystagmus in all directed toward the unchallenged ear (paralytic). The caloric response time courses were classified into two types, which were irreversible type and reversible type after endolymphatic sac KLH challenge. The immune injury of animals with irreversible type was thought to be stronger than that of these with reversible type. The spontaneous nystagmus of irreversible type animals was longer than that of reversible type animals. The temporary vestibular dysfunction was thought to be similar to that observed in Meniere's disease.

Intermediate Filaments in the Endolymphatic Sac of the Guinea Pig

The expression of the five main groups of intermediate filaments and their subgroups, especially cytokeratins, was investigated in the guinea pig endolymphatic sac at the light microscopic level using immunohistochemistry. Immunostaining for cytokeratin (PKK1, PKK2, PKK3) was found in the epithelial cell layer of the sac. Vimentin was seen to stain epithelial cell layer as well as subepithelial tissue. These findings may indicate that the cytokeratins are closely related to the inner ear fluid transport. The coexpression of cytokeratin and vimentin seemed to indicate the dual function of absorption and secretion of the endolymphatic sac epithelial cell to regulate the inner ear fluid homeostasis.

Lectin detection of carbohydrates in the endolymphatic sac.

The carbohydrate contents of the guinea pig endolymphatic sac were investigated by the use of lectins. The lumen of the endolymphatic sac was filled with stainable precipitate containing N-acetyl glucosamine, mannose, glucose, galactose and fucose. N-Acetyl galactosamine was also detected but in minute amounts. This composition corresponded to other areas in the inner ear, such as the cupula, the otolithic membrane and the tectorial membrane. The function of these carbohydrates may play an important role in preventing the lumen of the endolymphatic sac from collapsing as well as in regulating transepithelial fluid transport.

The Phagocytotic Activity of the Endolymphatic Sac in Guinea Pigs

The Phagocytotic Activity of the Endolymphatic Sac in Guinea Pigs

An Anionic Charge Barrier in the Guinea Pig Endolymphatic Sac

An Anionic Charge Barrier in the Guinea Pig Endolymphatic Sac

モルモット内リンパ嚢における中間径フィラメントの局在について

The expression of the five main groups of intermediate filaments and their subgroups, especially cytokeratins, was investigated in the guinea pig endolymphatic sac by light microscopy and immunohistochemistry. Immunoreaction for cytokeratin (PKK1, PKK2, PKK3) was found in the epithelial cell layer of the sac. Vimentin was seen to stain the epithelial cell layer as well as subepithelial tissue. These findings suggest that the cytokeratins are involved in the inner ear fluid transporting mechanism. The co-expression of cytokeratin and vimentin may be closely related to the dynamic properties of the endolymphatic sac which regulate inner ear fluid homeostasis.

Ruthenium red staining of the endolymphatic sac in the guinea pig

The ultrastructure of the guinea pig endolymphatic sac was studied, using the ruthenium red staining technique. The dye stained the apical cell surface coat and the homogeneous substance in the luminal space of the endolymphatic sac, when introduced from the luminal side of the epithelium. It is suggested that the surface coat and homogeneous substance may play an important part in the endolymph regulatory mechanism in the endolymphatic sac. When ruthenium red was introduced from the subepithelial side, the basolateral surface of the epithelial cells usually became brightly stained in the absence of staining of the apical cell surface, due to the presence of the tight junctions. In some instances, however, the dye penetrated beyond the level of the tight junctions. Pinocytotic vesicles and larger vacuoles in the epithelial cells were also sometimes stained, both apically and near the lateral cell surface. These findings suggest that endolymph efflux mechanisms in the endolymphatic sac may involve the combined actions of a paracellular and transepithelial flow as well as a transcellular, vacuolar bulk flow.

Cytochemical localization of Na-K ATPase in the guinea pig endolymphatic sac.

The light microscopical and ultracytochemical localization of Na-K ATPase (ouabain-sensitive, potassium-dependent p-nitrophenylphosphatase) in the guinea pig endolymphatic sac was studied by a newly developed lead citrate one step method. The cytochemical reaction product was detected in the epithelial cell layer of the endolymphatic sac at the light microscopical level. At the electron microscopical level, the reaction product was located to the basolateral plasma membrane of both the dark and the light cells. The results suggest that Na-K ATPase may play a role in the ion transport mechanism in the endolymphatic sac.

Ultrastructural localization of carbonic anhydrase and its possible role in the endolymphatic sac.

Carbonic anhydrase was detected histochemically in the guinea pig endolymphatic sac. The enzyme reaction was positive in both light and dark epithelial cells. In the former, the reaction product was found in the cytoplasm, especially around the intracytoplasmic vesicles and vacuoles. Reaction product was found in the basolateral infoldings as well. The dark epithelial cells also displayed carbonic anhydrase activity in the cytoplasm as well as in the lysosome-like bodies. It is suggested that this enzyme may be involved in (1) ionic or fluid regulation of the endolymph, (2) otoconia metabolism, and (3) phagocytotic activities.

Three-dimensional ultrastructure of the endolymphatic sac.

The subcellular structures of the epithelial cells of the guinea pig endolymphatic sac were studied. By using a newly developed scanning electron microscopy technique, the intracellular organelles could be studied three-dimensionally and the topographic relationships analyzed. The light epithelial cell has an extensive network of endoplasmic reticulum which is characteristically arranged in a baso-apical direction. The connections between the inner surface of the plasmalemma and the endoplasmic reticulum were observed, as were connections between the Golgi complex and the endoplasmic reticulum. Our findings support the hypothesis that the endoplasmic reticulum might form transcellular channels through which the cell can transport water and ions from the lumen of the endolymphatic sac out into the subepithelial tissue. The dark epithelial cells seen in particular contained the smooth type of endoplasmic reticulum. Lysosomes were also observed in the dark cells, indicating that these cells probably have more of a secretory function.