Apoptosis in the developing zebrafish embryo.

Apoptosis is a major part of the normal development of many organ systems and tissues. The zebrafish (Danio rerio) has become a useful model for studying early development, and recent advances in techniques used to label apoptotic cells have made it possible to visualize apoptotic cells in this model system. We have used the in situ terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL) to describe the temporal and spatial distribution of apoptotic cells during normal development of the zebrafish embryo from 12 to 96 h postfertilization. By counting labeled apoptotic cells, we have demonstrated transient high rates of cell death in various structures during development, and we have correlated these peaks with previously described developmental changes in these structures. Our analysis has focused on the nervous system and associated sensory organs including the olfactory organ, retina, lens, cornea, otic vesicle, lateral line organs, and Rohon-Beard neurons. Apoptosis is also described in other non-neural structures such as the notochord, somites, muscle, tailbud, and fins.

Patterning of the mammalian cochlea.

The mammalian cochlea is sophisticated in its function and highly organized in its structure. Although the anatomy of this sense organ has been well documented, the molecular mechanisms underlying its development have remained elusive. Information generated from mutant and knockout mice in recent years has increased our understanding of cochlear development and physiology. This article discusses factors important for the development of the inner ear and summarizes cochlear phenotypes of mutant and knockout mice, particularly Otx and Otx2. We also present data on gross development of the mouse cochlea.

Sensory organ generation in the chicken inner ear: contributions of bone morphogenetic protein 4, serrate1, and lunatic fringe.

The chicken inner ear is a remarkably complex structure consisting of eight morphologically distinct sensory organs. Unraveling how these sensory organs are specified during development is key to understanding how such a complex structure is generated. Previously, we have shown that each sensory organ in the chicken inner ear arises independently in the rudimentary otocyst based on Bone morphogenetic protein 4 (Bmp4) expression. Here, we compare the expression of Bmp4 with two other putative sensory organ markers, Lunatic Fringe (L-fng) and chicken Serrate1 (Ser1), both of which are components of the Notch signaling pathway. L-fng and Ser1 expression domains were asymmetrically distributed in the otic cup. At this early stage, expression of L-fng is similar to Delta1 (Dl1), in an anteroventral domain apparently corresponding to the neurogenic region, while Ser1 is expressed at both the anterior and posterior poles. By the otocyst stage, the expression of both L-fng and Ser1 largely coincided in the medial region. All presumptive sensory organs, as identified by Bmp4 expression, arose within the broad L-fng- and Ser1-positive domain, indicating the existence of a sensory-competent region in the rudimentary otocyst. In addition, there is a qualitative difference in the levels of expression between L-fng and Ser1 such that L-fng expression was stronger in the ventral anterior, whereas Ser1 was stronger in the dorsal posterior region of this broad domain. This early difference in expression may presage the differences among sensory organs as they arise from this sensory competent zone.

Impedance audiometric measurements in clinically normal dogs.

OBJECTIVE: To measure impedance audiometric values in clinically normal dogs that were sedated or anesthetized, evaluate effects of ear flushing on tympanometric measurements, and determine effects of performing acoustic reflex testing in a sound-attenuated room. ANIMALS: 35 mixed-breed and purebred client-owned dogs and 21 laboratory-bred Beagles. PROCEDURES: Tympanometry and acoustic reflex testing were performed on 27 mixed-breed and purebred dogs under isoflurane anesthesia in a non-sound-attenuated room and 21 Beagles under sedation in a sound-attenuated room. Tympanometry was performed on 8 mixed-breed dogs under halothane anesthesia before and after ear canal flushing. RESULTS: Among impedance audiometric values, ear canal volume and compliance peak were smaller in Beagles than in mixed-breed dogs; differences among other values were not detected. Ear canal volume was dependent on body weight. Differences were not found for tympanometric values measured before and after ear canal flushing. CONCLUSIONS AND CLINICAL RELEVANCE: Results of this study established reference range values for impedance audiometric measurements in clinically normal dogs under isoflurane anesthesia or sedation. Acoustic reflex testing does not need to be performed in a sound-attenuated room. The ear canals of clinically normal dogs can be flushed prior to performing tympanometry without altering the results. Impedance audiometry may be a useful noninvasive procedure for the diagnosis of otitis media in dogs.

Microbial flora and antimicrobial susceptibility patterns of isolated pathogens from the horizontal ear canal and middle ear in dogs with otitis media.

OBJECTIVE: To compare microbial flora and antimicrobial susceptibility patterns of isolated pathogens from the horizontal ear canal and middle ear in dogs with otitis media. DESIGN: Prospective study. ANIMALS: 23 dogs with chronic bilateral otitis externa. PROCEDURES: Swab specimens of the horizontal ear canal and middle ear were obtained for cytologic analysis, bacterial culture, and antimicrobial susceptibility testing. Integrity of the tympanic membrane was observed. If the tympanic membrane was intact, myringotomy was performed to collect specimens. RESULTS: Otitis media was diagnosed in 38 of 46 (82.6%) ears evaluated. The tympanic membrane was intact in 71.1% of the ears with otitis media. The 3 most common organisms isolated from the horizontal ear canal and middle ear were Staphylococcus intermedius, yeast, and Pseudomonas spp. A difference in total isolates or susceptibility patterns between the horizontal ear canal and middle ear was found in 34 (89.5%) ears. Compared with results of bacterial culture, cytologic examination of swab specimens was not as effective for detection of rods and cocci from the middle ear. CLINICAL IMPLICATIONS: In dogs with chronic otitis externa, otitis media often exists even when there is an intact tympanic membrane. In our study, the same isolates were rarely found in the horizontal ear canal and middle ear. Therefore, to choose appropriate antimicrobial agents, in addition to cytologic examination, bacterial culture and susceptibility testing of swab specimens from the horizontal ear canal and middle ear should be performed.