Neuromuscular Junction Development Differs Between Extraocular and Skeletal Muscles and Between Different Extraocular Muscles.

Purpose: To determine whether development of neuromuscular junctions (NMJs) differs between extraocular muscles (EOMs) and other skeletal muscles. Methods: Mouse EOMs, diaphragm, and tibialis anterior (TA) were collected at postnatal day (P)0, P3, P7, P10, P14, and P21, and 12 weeks. Whole muscles were stained with α-bungarotoxin, anti-neurofilament antibody, and slow or fast myosin heavy chain antibody, and imaged with a confocal microscope. Images were quantified using Imaris software. Results: NMJs in the EOMs show a unique pattern of morphological development compared to diaphragm and TA. At P0, diaphragm and TA NMJs were oval plaques; EOM single NMJs were long, thin rods. NMJs in the three muscle types progress to mature morphology at different rates. At all ages, EOM single NMJs were larger, especially relative to myofiber size. The inferior oblique and inferior rectus muscles show delayed single NMJ development compared to other EOMs. NMJs on multiply-innervated fibers in the EOMs vary widely in size, and there were no consistent differences between muscles or over time. Incoming motor nerves formed complex branching patterns, dividing first into superficial and deep branches, each of which branched extensively over the full width of the muscle. Motor axons that innervate multiply-innervated fibers entered the muscle with the axons that innervate singly-innervated fibers, then extended both proximally and distally. EOM NMJs had more subsynaptic nuclei than skeletal muscle NMJs throughout development. Conclusions: EOMs show a unique pattern of NMJ development and have more subsynaptic nuclei than other muscles, which may contribute to the exquisite control of eye movements.

Oculomotor nerve guidance and terminal branching requires interactions with differentiating extraocular muscles.

Muscle function is dependent on innervation by the correct motor nerves. Motor nerves are composed of motor axons which extend through peripheral tissues as a compact bundle, then diverge to create terminal nerve branches to specific muscle targets. As motor nerves approach their targets, they undergo a transition where the fasciculated nerve halts further growth then after a pause, the nerve later initiates branching to muscles. This transition point is potentially an intermediate target or guidepost to present specific cellular and molecular signals for navigation. Here we describe the navigation of the oculomotor nerve and its association with developing muscles in mouse embryos. We found that the oculomotor nerve initially grew to the eye three days prior to the appearance of any extraocular muscles. The oculomotor axons spread to form a plexus within a mass of cells, which included precursors of extraocular muscles and other orbital tissues and expressed the transcription factor Pitx2. The nerve growth paused in the plexus for more than two days, persisting during primary extraocular myogenesis, with a subsequent phase in which the nerve branched out to specific muscles. To test the functional significance of the nerve contact with Pitx2+ cells in the plexus, we used two strategies to genetically ablate Pitx2+ cells or muscle precursors early in nerve development. The first strategy used Myf5-Cre-mediated expression of diphtheria toxin A to ablate muscle precursors, leading to loss of extraocular muscles. The oculomotor axons navigated to the eye to form the main nerve, but subsequently largely failed to initiate terminal branches. The second strategy studied Pitx2 homozygous mutants, which have early apoptosis of Pitx2-expressing precursor cells, including precursors for extraocular muscles and other orbital tissues. Oculomotor nerve fibers also grew to the eye, but failed to stop to form the plexus, instead grew long ectopic projections. These results show that neither Pitx2 function nor Myf5-expressing cells are required for oculomotor nerve navigation to the eye. However, Pitx2 function is required for oculomotor axons to pause growth in the plexus, while Myf5-expressing cells are required for terminal branch initiation.

Local retinoic acid signaling directs emergence of the extraocular muscle functional unit.

Coordinated development of muscles, tendons, and their attachment sites ensures emergence of functional musculoskeletal units that are adapted to diverse anatomical demands among different species. How these different tissues are patterned and functionally assembled during embryogenesis is poorly understood. Here, we investigated the morphogenesis of extraocular muscles (EOMs), an evolutionary conserved cranial muscle group that is crucial for the coordinated movement of the eyeballs and for visual acuity. By means of lineage analysis, we redefined the cellular origins of periocular connective tissues interacting with the EOMs, which do not arise exclusively from neural crest mesenchyme as previously thought. Using 3D imaging approaches, we established an integrative blueprint for the EOM functional unit. By doing so, we identified a developmental time window in which individual EOMs emerge from a unique muscle anlage and establish insertions in the sclera, which sets these muscles apart from classical muscle-to-bone type of insertions. Further, we demonstrate that the eyeballs are a source of diffusible all-trans retinoic acid (ATRA) that allow their targeting by the EOMs in a temporal and dose-dependent manner. Using genetically modified mice and inhibitor treatments, we find that endogenous local variations in the concentration of retinoids contribute to the establishment of tendon condensations and attachment sites that precede the initiation of muscle patterning. Collectively, our results highlight how global and site-specific programs are deployed for the assembly of muscle functional units with precise definition of muscle shapes and topographical wiring of their tendon attachments.

Zebrafish dscaml1 Deficiency Impairs Retinal Patterning and Oculomotor Function.

Down syndrome cell adhesion molecules (dscam and dscaml1) are essential regulators of neural circuit assembly, but their roles in vertebrate neural circuit function are still mostly unexplored. We investigated the functional consequences of dscaml1 deficiency in the larval zebrafish (sexually undifferentiated) oculomotor system, where behavior, circuit function, and neuronal activity can be precisely quantified. Genetic perturbation of dscaml1 resulted in deficits in retinal patterning and light adaptation, consistent with its known roles in mammals. Oculomotor analyses revealed specific deficits related to the dscaml1 mutation, including severe fatigue during gaze stabilization, reduced saccade amplitude and velocity in the light, greater disconjugacy, and impaired fixation. Two-photon calcium imaging of abducens neurons in control and dscaml1 mutant animals confirmed deficits in saccade-command signals (indicative of an impairment in the saccadic premotor pathway), whereas abducens activation by the pretectum-vestibular pathway was not affected. Together, we show that loss of dscaml1 resulted in impairments in specific oculomotor circuits, providing a new animal model to investigate the development of oculomotor premotor pathways and their associated human ocular disorders.SIGNIFICANCE STATEMENTDscaml1 is a neural developmental gene with unknown behavioral significance. Using the zebrafish model, this study shows that dscaml1 mutants have a host of oculomotor (eye movement) deficits. Notably, the oculomotor phenotypes in dscaml1 mutants are reminiscent of human ocular motor apraxia, a neurodevelopmental disorder characterized by reduced saccade amplitude and gaze stabilization deficits. Population-level recording of neuronal activity further revealed potential subcircuit-specific requirements for dscaml1 during oculomotor behavior. These findings underscore the importance of dscaml1 in the development of visuomotor function and characterize a new model to investigate potential circuit deficits underlying human oculomotor disorders.

Morphology of the extraocular muscles (m. bulbi) in the pre-hatchling and post-hatchling african black ostriches (struthio camelus domesticus L., 1758) (Aves: Struthioniformes).

The aim of the study was to describe the morphology and the development of the extraocular muscles (EOMs) in the pre-hatchling and post-hatchling African black ostrich. The study involved 50 birds aged between 28 days and 3 years. The EOMs were analyzed morphologically with respect to the location and length of the straight and oblique muscles and the third eyelid muscles, the length and breadth of their tendons as well as the distance and shape of the muscle tendon insertions at the corneal limbus. A histological and histometric analysis were also carried out. The greatest increase in the length of the EOMs was noted in groups III-V. A marked increase in the length of the tendons of the dorsal straight muscle was found in groups II and III, in the tendons of the nasal straight muscle in groups IV and V, in the tendons of the dorsal oblique muscle in groups III to V and in the tendons of the ventral oblique muscle in groups IV and V. There was a significant increase in the breadth of the dorsal straight and ventral oblique muscle tendons in groups IV and V and the tendons of the pyramidal muscle in groups III and V. The distance of the distal insertion of the tendon at the corneal limbus increased steadily with age in all the examined groups. The number of fascicles and muscle fibres, their diameter and length in all the studied EOMs were different in the different groups.

Sparing of the extraocular muscles in mdx mice with absent or reduced utrophin expression: A life span analysis.

Sparing of the extraocular muscles in muscular dystrophy is controversial. To address the potential role of utrophin in this sparing, mdx:utrophin(+/-) and mdx:utrophin(-/-) mice were examined for changes in myofiber size, central nucleation, and Pax7-positive and MyoD-positive cell density at intervals over their life span. Known to be spared in the mdx mouse, and contrary to previous reports, the extraocular muscles from both the mdx:utrophin(+/-) and mdx:utrophin(-/-) mice were also morphologically spared. In the mdx:utrophin(+/)(-) mice, which have a normal life span compared to the mdx:utrophin(-/-) mice, the myofibers were larger at 3 and 12 months than the wild type age-matched eye muscles. While there was a significant increase in central nucleation in the extraocular muscles from all mdx:utrophin(+/)(-) mice, the levels were still very low compared to age-matched limb skeletal muscles. Pax7- and MyoD-positive myogenic precursor cell populations were retained and were similar to age-matched wild type controls. These results support the hypothesis that utrophin is not involved in extraocular muscle sparing in these genotypes. In addition, it appears that these muscles retain the myogenic precursors that would allow them to maintain their regenerative capacity and normal morphology over a lifetime even in these more severe models of muscular dystrophy.

Monocular and binocular development in children with albinism, infantile nystagmus syndrome, and normal vision.

BACKGROUND/AIMS: To compare interocular acuity differences, crowding ratios, and binocular summation ratios in 4- to 8-year-old children with albinism (n = 16), children with infantile nystagmus syndrome (n = 10), and children with normal vision (n = 72). METHODS: Interocular acuity differences and binocular summation ratios were compared between groups. Crowding ratios were calculated by dividing the single Landolt C decimal acuity with the crowded Landolt C decimal acuity mono- and binocularly. A linear regression analysis was conducted to investigate the contribution of 5 predictors to the monocular and binocular crowding ratio: nystagmus amplitude, nystagmus frequency, strabismus, astigmatism, and anisometropia. RESULTS: Crowding ratios were higher under mono- and binocular viewing conditions for children with infantile nystagmus syndrome than for children with normal vision. Children with albinism showed higher crowding ratios in their poorer eye and under binocular viewing conditions than children with normal vision. Children with albinism and children with infantile nystagmus syndrome showed larger interocular acuity differences than children with normal vision (0.1 logMAR in our clinical groups and 0.0 logMAR in children with normal vision). Binocular summation ratios did not differ between groups. Strabismus and nystagmus amplitude predicted the crowding ratio in the poorer eye (p = 0.015 and p = 0.005, respectively). The crowding ratio in the better eye showed a marginally significant relation with nystagmus frequency and depth of anisometropia (p = 0.082 and p = 0.070, respectively). The binocular crowding ratio was not predicted by any of the variables. CONCLUSIONS: Children with albinism and children with infantile nystagmus syndrome show larger interocular acuity differences than children with normal vision. Strabismus and nystagmus amplitude are significant predictors of the crowding ratio in the poorer eye.

A role for the macular carotenoids in visual motor response.

OBJECTIVES: Lutein (L), zeaxanthin (Z), and meso-zeaxanthin are the dominant carotenoids within the central retina (there, termed macular pigment, MP). L is also the dominant carotenoid in the brain. The presence of L and Z in both motor and visual areas of the central nervous system is consistent with a role of these carotenoids in visual-motor behavior. The purpose of this study was to provide a first test of this hypothesis. METHODS: Balance ability (measured via the Standing Leg Test) and simple reaction time (measured via a stimulus appearing in one of four quadrants of a computer monitor) were measured in 49 subjects (mean age = 54.8 years). Fixed and variable reaction time, and coincidence anticipation ability (estimating the arrival of the stimulus at a target location moving at four velocities) were assessed in 106 younger subjects (mean age = 23 years) using a customized device. MP optical density was measured in all subjects via customized heterochromatic flicker photometry. RESULTS: MP optical density was significantly (P < 0.05) related to reaction time and to balance ability for the older subjects. Even for the younger group, MP optical density was significantly (P < 0.05) related to fixed and variable position reaction time, as well as coincidence anticipation errors, at high speed. DISCUSSION: L and Z status has been linked to benefits in cognitive function in past research. The present results, and the selective presence of L and Z in visual and motor areas in the brain, are consistent with these carotenoids having a role in visual and motor integration.

Development transitions of thin filament proteins in rat extraocular muscles.

Extraocular muscles are a unique subset of striated muscles. During postnatal development, the extraocular muscles undergo a number of myosin isoform transitions that occur between postnatal day P10 (P10) and P15. These include: (1) loss of embryonic myosin from the global layer resulting in the expression restricted to the orbital layer; (2) the onset of expression of extraocular myosin and the putative tonic myosin (myh 7b/14); and (3) the redistribution of nonmuscle myosin IIB from a subsarcolemmal position to a sarcomeric distribution in the slow fibers of the global layer. For this study, we examined the postnatal appearance and distribution of α-actinin, tropomyosin, and nebulin isoforms during postnatal development of the rat extraocular muscles. Although sarcomeric α-actinin is detectable from birth, α-actinin 3 appears around P15. Both tropomyosin-1 and -2 are present from birth in the same distribution as in the adult animal. The expression of nebulin was monitored by gel electrophoresis and western blots. At P5-10, nebulin exhibits a lower molecular mass than observed P15 and later during postnatal development. The changes in α-actinin 3 and nebulin expression between P10 and P15 coincide with transitions in myosin isoforms as detailed above. These data point to P10-P15 as the critical period for the maturation of the extraocular muscles, coinciding with eyelid opening.

[Surgical management of congenital eyelid ptosis with silicone rod--outcome analysis]. / Ocena wyników leczenia operacyjnego wrodzonego opadniecia powieki górnej z zastosowaniem ekspandera silikonowego.

PURPOSE: To analyze the outcomes of surgical management of congenital eyelid ptosis with the use of silicone rod. MATERIAL AND METHODS: Congenital eyelid ptosis was diagnosed in 12 children aged 2-18 years: 6 cases with unilateral, and 6 with bilateral ptosis. Analyzing the patients we focused on the function of levator muscle before and after the surgery in particular. Ptosis repair was performed with the use of silicone rod in all children. RESULTS: Functional and cosmetic outcomes of the procedure with the silicone rod were good.

Genomic profiling reveals Pitx2 controls expression of mature extraocular muscle contraction-related genes.

PURPOSE: To assess the influence of the Pitx2 transcription factor on the global gene expression profile of extraocular muscle (EOM) of mice. METHODS: Mice with a conditional knockout of Pitx2, designated Pitx2(Δflox/Δflox) and their control littermates Pitx2(flox/flox), were used. RNA was isolated from EOM obtained at 3, 6, and 12 weeks of age and processed for microarray-based profiling. Pairwise comparisons were performed between mice of the same age and differentially expressed gene lists were generated. Select genes from the profile were validated using real-time quantitative polymerase chain reaction and protein immunoblot. Ultrastructural analysis was performed to evaluate EOM sarcomeric structure. RESULTS: The number of differentially expressed genes was relatively small. Eleven upregulated and 23 downregulated transcripts were identified common to all three age groups in the Pitx2-deficient extraocular muscle compared with littermate controls. These fell into a range of categories including muscle-specific structural genes, transcription factors, and ion channels. The differentially expressed genes were primarily related to muscle contraction. We verified by protein and ultrastructural analysis that myomesin 2 was expressed in the Pitx2-deficient mice, and this was associated with development of M lines evident in their orbital region. CONCLUSIONS: The global transcript expression analysis uncovered that Pitx2 primarily regulates a relatively select number of genes associated with muscle contraction. Pitx2 loss led to the development of M line structures, a feature more typical of other skeletal muscle.

Postnatal changes in the developing rat extraocular muscles.

PURPOSE: To examine the distribution and timing of expression of nonmuscle myosin IIB (nmMyH IIB) and the extraocular muscle (EOM)-specific myosin (EO-MyHC) during postnatal development of the rat extraocular muscles. METHODS: Whole orbits were collected from postnatal development day (P) 1 through P30 from Sprague-Dawley rats. Samples were analyzed by immunofluorescence microscopy and Western blot to examine the distribution and abundance of nmMyH IIB and EO-MyHC compared with other myosin isoforms and sarcomeric α-actinin. Polyclonal antibodies were produced to specifically detect EO-MyHC. Postnatal limb muscles were used as control. RESULTS: Analysis of EOM morphology in the developing orbits indicates that the global and orbital layers are not evident until day P15. The distribution of nmMyH IIB changes between days P10 and P15 from a subsarcolemma distribution to an intrafiber distribution in the global layer. EO-MyHC appears by day p15, primarily in the orbital layer of the EOMs. Sarcomeric α-actinin was equally abundant in the EOMs at all stages. Fetal MyHC was the predominant isoform at day P1 but slowly diminished in abundance with age in a layer-specific manner. CONCLUSIONS: These data demonstrate that significant changes occur in the EOMs from P10 to P15 and suggest that visual stimulation may play a role in the signals that regulate both nmMyH IIB and EO-MyHC developmental transitions. The pronounced distinctions of the orbital and global layers occurring by P15 establish the adult morphologic phenotype of the muscle.

Myosin heavy chain expression in mouse extraocular muscle: more complex than expected.

PURPOSE: To characterize the expression patterns of myosin heavy chain (MyHC) isoforms in mouse extraocular muscles (EOMs) during postnatal development. METHODS: MyHC isoform expression in mouse EOMs from postnatal day (P)0 to 3 months was evaluated by quantitative polymerase chair reaction (qPCR) and immunohistochemistry. The longitudinal and cross-sectional distribution of each MyHC isoform and coexpression of certain isoforms in single muscle fibers was determined by single, double, and triple immunohistochemistry. RESULTS: MyHC isoform expression in postnatal EOMs followed the developmental rules observed in other skeletal muscles; however, important exceptions were found. First, developmental isoforms were retained in the orbital layer of the adult EOMs. Second, expression of emb-MyHC, neo-MyHC, and 2A-MyHC was restricted to the orbital layer and that of 2B-MyHC to the global layer. Third, although slow-MyHC and 2B-MyHC did not exhibit obvious longitudinal variations, emb-MyHC, neo-MyHC, and 2A-MyHC were more abundant distally and were excluded from the innervational zone, whereas eom-MyHC complemented their expression and was more abundant in the mid-belly region in both the orbital and global layers. Fourth, coexpression of MyHC isoforms in single global layer fibers was rare, but it was common among the orbital layer fibers. CONCLUSIONS: MyHC isoforms have complex expression patterns, exhibiting not only longitudinal and cross-sectional variation of each isoform, but also of coexpression in single fibers. The highly heterogeneous MyHC expression reflects the complex contractile profiles of EOMs, which in turn are a function of the requirements of eye movements, which range from extremely fast saccades to sustained position, each with a need for precise coordination of each eye.

Schwann cells as a source of insulin-like growth factor-1 for extraocular muscles.

Precise force regulation is fundamentally important for extraocular muscle (EOM) function. Insulin-like growth factor-1 (IGF-1) plays a major role in EOM force regulation, but the source of endogenous IGF-1 is unclear. Multiple IGF-1 sources may supply EOMs, including: the EOM itself; the systemic circulation; innervating motoneurons; and Schwann cells within nerves. IGF-1 expression was measured in chicken during oculomotor system maturation by using real-time polymerase chain reaction (PCR). Accumulation of radiolabeled IGF-1 in EOMs was compared after either injection into the vascular circulation or into the trochlear nerve. Schwann cells were the most prominent IGF-1 source. A microtubule-dependent mechanism exists to anterogradely transport IGF-1 to EOMs. EOMs were significantly more efficient in extracting IGF-1 from the nerve than from the systemic circulation. Therefore, Schwann cells are the most prominent and potentially the most important source of IGF-1 for EOMs. These findings may contribute to a better understanding of EOM force regulation and its failure in strabismus.

A Comparação de funções apendiculares desencadeadas pela visão em lactentes nascidos pré-termo e a termo no primeiro trimestre de vida / Comparison of appendicular functions released trough sight of pre-term newly born infants and to a term in the first 3 months of life

OBJETIVO: caracterizar e comparar o desenvolvimento de funções apendiculares entre dois grupos de lactentes a termo e pré-termo e verificar se a fixação visual pode ser considerada como um pré-requisito para as ações dos membros superiores. MÉTODO: estudo realizado no Centro de Estudos e Pesquisas em Reabilitação"Prof. Dr. Gabriel Porto", da Universidade Estadual de Campinas em dois grupos: 1) pré-termo, constituído de 21 lactentes, com idade corrigida entre 1 e 3 meses e 2) 21 recém-nascido a termo. Para avaliação utilizou-se o Método de Avaliação da Conduta Visual de Lactentes. Na análise estatística utilizou-se o teste "Q"de Cochran e o teste de Yates. RESULTADOS: verificou-se que, embora, no primeiro mês de vida a fixação visual tenha sido mais frequente no grupo a termo, não se observaram valores de significância estatística para cada grupo e entre eles. Verificou-se diferença estatística mês a mês para o grupo pré-termo e a termo em ambas as funções apendiculares, observando-se o mesmo valor de significância estatística (p = 0,000). Na comparação entre os grupos, tais funções não mostraram diferença significativa, embora tenha sido observado menores frequências nos pré-termo. Apesar da sequência das aquisições apendiculares desencadeadas pela visão ser semelhante nos grupos, o ritmo e o padrão de desenvolvimento apendicular dos lactentes pré-termo, com a idade corrigida, foram diferentes dos a termo, nos aspectos de frequência e qualidade

An altered phenotype in a conditional knockout of Pitx2 in extraocular muscle.

PURPOSE: To determine the temporal and spatial expression of Pitx2, a bicoid-like homeobox transcription factor, during postnatal development of mouse extraocular muscle and to evaluate its role in the growth and phenotypic maintenance of postnatal extraocular muscle. METHODS: Mouse extraocular muscles of different ages were examined for the expression of Pitx2 by RT-PCR, q-PCR, and immunostaining. A conditional mutant mouse strain, in which Pitx2 function is inactivated at postnatal day (P)0, was generated with a Cre-loxP strategy. Histology, immunostaining, real-time PCR, in vitro muscle contractility, and in vivo ocular motility were used to study the effect of Pitx2 depletion on extraocular muscle. RESULTS: All three Pitx2 isoforms were expressed by extraocular muscle and at higher levels than in other striated muscles. Immunostaining demonstrated the presence of Pitx2 mainly in extraocular muscle myonuclei. However, no obvious expression patterns were observed in terms of anatomic region (orbital versus global layer), innervation zone, or muscle fiber types. The mutant extraocular muscle had no obvious pathology but had altered muscle fiber sizes. Expression levels of myosin isoforms Myh1, Myh6, Myh7, and Myh13 were reduced, whereas Myh2, Myh3, Myh4, and Myh8 were not affected by postnatal loss of Pitx2. In vitro, Pitx2 loss made the extraocular muscles stronger, faster, and more fatigable. Eye movement recordings found saccades to have a lower peak velocity. CONCLUSIONS: Pitx2 is important in maintaining the mature extraocular muscle phenotype and regulating the expression of critical contractile proteins. Modulation of Pitx2 expression can influence extraocular muscle function with long-term therapeutic implications.

Classification and development of myofiber types in the superior oblique extraocular muscle of chicken.

Precise control of contractile force of extraocular muscles is required for appropriate movements and alignment of the eyes. It is unclear how such precise regulation of contractile force is achieved during development and maturation. By using the posthatch chicken as a model, we describe and quantify critical parameters of the developing superior oblique extraocular muscle from hatching to 16 weeks of age, including contractile force, muscle mass, myofiber diameters, classification of fiber types, and distribution and quantification of mitochondria. Analysis at the light- and electron microscopic levels shows that chicken myofiber types largely correspond to their mammalian counterparts, with four fiber types in the orbital and four types in the global layer. Twitch tension muscle force and muscle mass gradually increase and stabilize at approximately 11 weeks. Tetanic tension continues to increase between 11 and 16 weeks. Myofiber diameters in both the orbital and global layer increase from hatching to six weeks, and then stabilize, whereas the myofiber number is constant after hatching. This finding suggests that muscle mass increases during late maturation due to increasing fiber length rather than fiber diameter. Quantitative ultrastructural analysis reveals continuing changes in the composition of the four muscle fiber types, suggesting ongoing fiber type conversion or differential replacement of myofiber types. Muscle fiber composition continues to change into late juvenile and adult age. Our study provides evidence for gradual, incremental, and continuing changes in avian myofiber composition and function that is similar to postnatal oculomotor maturation in visually oriented mammals such as kitten.

[Variation of nestin expression in SD rat eyes in different stages of postnatal development].

OBJECTIVE: To observe the position and quantity of nestin expression in SD rat eyes in different stages of postnatal development. METHODS: Immunocytochemical method was used to identify nestin expression in the eyes of SD rats of 1 to 30 days old. RESULTS: Nestin expression was detected in the retina and extraocular muscles of SD rats. The expression varied with the time of postnatal development, distributing in the entire retina layers in earlier stages and confined in the nerve fiber layer in later stages. The quantities of nestin expression in the extraocular muscles decreased gradually with growth. CONCLUSION: Nestin expression in the retinas and extraocular muscles of SD rats decreases during the postnatal development.

Extraocular muscle quantification using mathematical morphology: a semi-automatic method for analyzing muscle enlargement in orbital diseases.

Mathematical morphology was used to quantify the dimensions of the human extraocular muscles in computed tomography orbital scans. Coronal images of 28 patients with Graves orbitopathy and 5 controls (218 orbital scans) were analyzed with an algorithm for extraocular muscle segmentation. The results showed that measurements of extraocular muscle area obtained with semi-automatic segmentation are highly correlated with manual tracing and provides a simple method to quantify orbital structures including the extraocular muscles.

Postnatal development of myosin heavy chain isoforms in rat extraocular muscles.

PURPOSE: To determine the composition of myosin heavy chain (MHC) isoforms in rat extraocular muscles (EOMs) during postnatal development. METHODS: The MHC composition of rat EOMs at postnatal day 0 (P0), postnatal day 14 (P14), and adults was evaluated at mRNA levels by competitive polymerase chain reaction and MHC composition of each six EOM in adult rats. RESULTS: EOMs at P0 revealed predominant expression of neonatal MHC (75.5%) with a lesser percentage of embryonic MHC (12.8%) and 2A MHC (11.5%). 2X MHC was expressed at low levels and other MHC isoforms were not detected. At P14, EOMs expressed mostly 2X MHC (42.4%) and 2A MHC (27.4%). Expression levels of neonatal MHC (14.1%) and embryonic MHC (4.9%) decreased. 2B MHC (8.2%), EOM MHC (1.9%), and beta-cardiac MHC (1.1%) were detected at low levels. In the adult rats, EOMs contained over 80% of three fast MHC isoforms, such as 2X MHC (29.9%), 2A MHC (29.3%), and 2B MHC (24.5%). Each of six adult EOM showed slightly different expression levels of MHC composition. CONCLUSIONS: A strong correlation exists between the composition of fast MHC isoforms and muscle development. MHC isoform followed a neonatal MHC-2X MHC-2B MHC isoform switching pattern after birth. Postnatal development of EOMs had a slightly different expression pattern for MHC isoforms and may have different regulatory roles related to their functional requirement.