Effects of Weightlessness on Vestibular Development of Quail

The lack of gravity is known to alter vestibular responses in developing and adult vertebrates. One cause of these altered responses may be changes in the connections between the vestibular receptor and the brain. Therefore we propose to investigate the effects of gravity on the formations of connections between the gravity receptors of the ear and the brain in developing quail incubated in space beginning at an age before these connections are established (incubation day three) until near the time of hatching, when they are to some extent functional. This investigation will make use of a novel technique, the diffusion of a lipophilic dye, DiI, in fixed tissue. This technique can thus be used to analyze the connections in specimens fixed in orbit, thus eliminating changes due to the earth's gravity. The evaluation of the data will enable us to detect gross deviations from normal patterns as well as detailed quantitative deviations

Effects of Weightlessness on Vestibular Development of Quail

The data confirm previous findings that quail embryos can, under proper circumstances, develop until hatching in microgravity. There were no gross abnormalities in the few ears of the late embryos (we received 3 ears at E14.5 and 4 ears at E16.5). Due to inadequate numbers of samples returned and their fully insufficient fixation, no conclusions could be reached that warrant any publications

Connecting ears to eye muscles: Evolution of a 'simple' reflex arc

Developmental and evolutionary data from vertebrates are beginning to elucidate the origin of the sensorimotor pathway that links gravity and motion detection to image-stabilizing eye movements--the vestibulo-ocular reflex (VOR). Conserved transcription factors coordinate the development of the vertebrate ear into three functional sensory compartments (graviception/translational linear acceleration, angular acceleration and sound perception). These sensory components connect to specific populations of vestibular and auditory projection neurons in the dorsal hindbrain through undetermined molecular mechanisms. In contrast, a molecular basis for the patterning of the vestibular projection neurons is beginning to emerge. These are organized through the actions of rostrocaudally and dorsoventrally restricted transcription factors into a 'hodological mosaic' within which coherent and largely segregated subgroups are specified to project to different targets in the spinal cord and brain stem. A specific set of these regionally diverse vestibular projection neurons functions as the central element that transforms vestibular sensory signals generated by active and passive head and body movements into motor output through the extraocular muscles. The large dynamic range of motion-related sensory signals requires an organization of VOR pathways as parallel, frequency-tuned, hierarchical connections from the sensory periphery to the motor output. We suggest that eyes, ears and functional connections subserving the VOR are vertebrate novelties that evolved into a functionally coherent motor control system in an almost stereotypic organization across vertebrate taxa

Brn3c null mutant mice show long-term, incomplete retention of some afferent inner ear innervation

BACKGROUND: Ears of Brn3c null mutants develop immature hair cells, identifiable only by certain molecular markers, and undergo apoptosis in neonates. This partial development of hair cells could lead to enough neurotrophin expression to sustain sensory neurons through embryonic development. We have therefore investigated in these mutants the patterns of innervation and of expression of known neurotrophins. RESULTS: At birth there is a limited expression of BDNF and NT-3 in the mutant sensory epithelia and DiI tracing shows no specific reduction of afferents or efferents that resembles neurotrophin null mutations. At postnatal day 7/8 (P7/8), innervation is severely reduced both qualitatively and quantitatively. 1% of myosin VIIa-positive immature hair cells are present in the mutant cochlea, concentrated in the base. Around 20% of immature hair cells exist in the mutant vestibular sensory epithelia. Despite more severe loss of hair cells (1% compared to 20%), the cochlea retains many more sensory neurons (46% compared to 15%) than vestibular epithelia. Even 6 months old mutant mice have some fibers to all vestibular sensory epithelia and many more to the cochlear apex which lacks MyoVIIa positive hair cells. Topologically organized central cochlea projections exist at least until P8, suggesting that functional hair cells are not required to establish such projections. CONCLUSION: The limited expression of neurotrophins in the cochlea of Brn3c null mice suffices to support many sensory neurons, particularly in the cochlea, until birth. The molecular nature of the long term survival of apical spiral neurons remains unclear

Cranial sensory neuron development in the absence of brain-derived neurotrophic factor in BDNF/Bax double null mice

AbstractTo investigate the role of brain-derived neurotrophic factor (BDNF) in differentiation of cranial sensory neurons in vivo, we analyzed development of nodose (NG), petrosal (PG), and vestibular (VG) ganglion cells in genetically engineered mice carrying null mutations in the genes encoding BDNF and the proapoptotic Bcl-2 homolog Bax. In bax −/− mutants, ganglion cell numbers were increased significantly compared to wild-type animals, indicating that naturally occurring cell death in these ganglia is regulated by Bax signaling. Analysis of bdnf −/− bax −/− mutants revealed that, although the Bax null mutation completely rescued cell loss in the absence of BDNF, it did not rescue the lethality of the BDNF null phenotype. Moreover, despite rescue of BDNF-dependent neurons by the bax null mutation, sensory target innervation was abnormal in double null mutants. Vagal sensory innervation to baroreceptor regions of the cardiac outflow tract was completely absent, and the density of vestibular sensory innervation to the cristae organs was markedly decreased, compared to wild-type controls. Moreover, vestibular afferents failed to selectively innervate their hair cell targets within the cristae organs in the double mutants. These innervation failures occurred despite successful navigation of sensory fibers to the peripheral field, demonstrating that BDNF is required locally for afferent ingrowth into target tissues. In addition, the bax null mutation failed to rescue expression of the dopaminergic phenotype in a subset of NG and PG neurons. These data demonstrate that BDNF signaling is required not only to support survival of cranial sensory neurons, but also to regulate local growth of afferent fibers into target tissues and, in some cells, transmitter phenotypic expression is required

Combining Lipophilic dye, in situ Hybridization, Immunohistochemistry, and Histology

Going beyond single gene function to cut deeper into gene regulatory networks requires multiple mutations combined in a single animal. Such analysis of two or more genes needs to be complemented with in situ hybridization of other genes, or immunohistochemistry of their proteins, both in whole mounted developing organs or sections for detailed resolution of the cellular and tissue expression alterations. Combining multiple gene alterations requires the use of cre or flipase to conditionally delete genes and avoid embryonic lethality. Required breeding schemes dramatically enhance effort and cost proportional to the number of genes mutated, with an outcome of very few animals with the full repertoire of genetic modifications desired. Amortizing the vast amount of effort and time to obtain these few precious specimens that are carrying multiple mutations necessitates tissue optimization. Moreover, investigating a single animal with multiple techniques makes it easier to correlate gene deletion defects with expression profiles. We have developed a technique to obtain a more thorough analysis of a given animal; with the ability to analyze several different histologically recognizable structures as well as gene and protein expression all from the same specimen in both whole mounted organs and sections. Although mice have been utilized to demonstrate the effectiveness of this technique it can be applied to a wide array of animals. To do this we combine lipophilic dye tracing, whole mount in situ hybridization, immunohistochemistry, and histology to extract the maximal possible amount of data

A comprehensive analysis of interleukin-4 receptor polymorphisms and their association with atopy and IgE regulation in childhood

Background: The interleukin (IL) 4/IL13 pathway is involved in the regulation of IgE production associated with atopic diseases. Numerous polymorphisms have been identified in the coding region of the IL4 receptor alpha chain (IL4Ra) and previous association studies have shown conflicting results. Based on their putative functional role, polymorphisms A148G, T1432C and A1652G, located in the coding region of IL4Ra, were selected for association and haplotype studies in a large German population sample (n = 1,120). Methods: Genotyping was performed using allele-specific PCR and restriction-enzyme-based assays. Haplotypes were estimated, and population-derived IgE percentiles (50% IgE >60 IU/ml, 66% IgE >115 IU/ml and 90% IgE >457 IU/ml) were calculated as outcome variables in a haplotype trend regression analysis. Results: In our population, only polymorphism T1432C showed a trend for a protective effect against atopic rhinitis ( odds ratio, OR: 0.52, 95% confidence interval, CI: 0.26 - 1.02, p = 0.05). When haplotypes were calculated, one haplotype was significantly associated with elevated serum IgE levels at the 50th percentile ( OR 1.60, 95% CI 1.08 - 2.37, p = 0.02). Conclusions: These data indicate that IL4Ra polymorphisms, although suggested to be functionally relevant by in vitro studies, have only a minor influence on IgE regulation in our large population sample. Copyright (C) 2004 S. Karger AG, Basel

Observations On The Shape Of The Lens In The Eye Of the Silver Lamprey, Ichthyomyzon Unicuspis

The shape of the lens in the eye of the silver lamprey, ichthyomyzon unicuspis was examined in live, frozen, and fixed material. Contrary to other reports, the lens was found to be nonspherical with a cone-shaped posterior. The egg-shaped lens, which contains horizontal sutures on both the anterior and posterior surfaces, is also asymmetric in the nasotemporal axis. Its equatorial diameter exceeds its axail diameter (thickness) and the radius of curvature of the lens in the dorsoventral axis is greater than the radius of curvature in the anterioposterior axis. The lens is surrounded by a thick basement membrane with the anterior lens surface covered by a single layer of cuboidal epithelial cells. Juxtaposed to the lens capsule is a dense layer of lens fibres, which stain more darkly and surround an ill-defined lens nucleus. The shape of the lens is discussed in relation to that in aquatic gnathostomes and compared with the putative multifocal lenses of some mesopelagic teleosts. It is also hypothesized that the previously reported active focussing ability of the lamprey eye may have been misinterpreted, owing to failure to take into account the nonspherical lens shape, and may reflect measurements taken of the eye and lens at different angles

Auditory Nomenclature: Combining Name Recognition With Anatomical Description

The inner ear and its two subsystems, the vestibular and the auditory system, exemplify how the identification of distinct cellular or anatomical elements ahead of elucidating their function, leads to a medley of anatomically defined and recognition oriented names that confused generations of students. Past attempts to clarify this unyielding nomenclature had incomplete success, as they could not yet generate an explanatory nomenclature. Building on these past efforts, we propose a somewhat revised nomenclature that keeps most of the past nomenclature as proposed and follows a simple rule: Anatomical and explanatory terms are combined followed, in brackets, by the name of the discoverer (see Table 1). For example, the “organ of Corti” will turn into the spiral auditory organ (of Corti). This revised nomenclature build as much as possible on existing terms that have explanatory value while keeping the recognition of discoverers alive to allow a transition for those used to the eponyms. Once implements, the proposed terminology should help future generations in learning the structure-function correlates of the ear more easily. To facilitate future understanding, leading genetic identifiers for a given structure have been added wherever possible

Pax2 and Pax8 cooperate in mouse inner ear morphogenesis and innervation

<p>Abstract</p> <p>Background</p> <p><it>Pax2;5;8 </it>transcription factors play diverse roles in vertebrate and invertebrate organogenesis, including the development of the inner ear. Past research has suggested various cochlear defects and some vestibular defects in <it>Pax2 </it>null mice but the details of the cochlear defects and the interaction with other <it>Pax </it>family members in ear development remain unclear.</p> <p>Results</p> <p>We show that <it>Pax2;8 </it>double null mice do not develop an ear past the otocyst stage and show little to no sensory as well as limited and transient neuronal development, thus indicating that these two family members are essential for overall ear morphogenesis and sustained neurosensory development. In support of functional redundancy between Pax proteins, <it>Pax2 </it>can be substituted by a <it>Pax5 </it>minigene, a gene normally not expressed in the embryonic mouse ear. There is no detectable morphological defect in <it>Pax8 </it>null mice suggesting that <it>Pax2 </it>expression can compensate for <it>Pax8</it>. Conversely, <it>Pax8 </it>cannot compensate for <it>Pax2 </it>leading to a cochlear phenotype not fully appreciated previously: Cochlear development is delayed until E15.5 when the cochlea extrudes as a large sack into the brain case. Immunocytochemistry and tracing from the brain show that a cochlear spiral ganglia form as a small addition to the inferior vestibular ganglion. However, the empty cochlear sack, devoid of any sensory epithelium development as indicated by the absence of Sox2 or MyoVII expression, nevertheless develop a dense innervation network of small neurons situated in the wall of the cochlear sack.</p> <p>Conclusions</p> <p>Combined these data suggest that <it>Pax2 </it>is needed for organ of Corti formation and is directly or indirectly involved in the coordination of spiral ganglion formation which is partially disrupted in the <it>Pax2 </it>null ears. All three <it>Pax </it>genes can signal redundantly in the ear with their function being determined primarily by the spatio-temporal expression driven by the three distinct promoters of these genes.</p