Cranial Cartilage Research Articles

Identification of a protein in squid cranial cartilage with link protein properties

Squid cranial cartilage extracts were found to contain a protein with a molecular mass of 35 kDa immunoreacting with an antiserum against sheep link protein. Because hyaluronan is not detected in this tissue and the structure of proteoglycans is different to that of aggreean or versican, this observation was studied further. The 35 kDa protein was purified from cartilage extracts and immunolocalised in Western blots by both the polyclonal antibody and the mAb 8A4. It was found that it was able to bind to hyaluronan and to aggrecan. Direct and competitive microplate binding experiments showed that the squid protein binds to G1 domain of aggrecan, similarly to cartilage link protein and, therefore, it could be a link-like protein molecule of squid cranial cartilage. The 35 kDa protein was also able to bind to squid proteoglycan and this suggested that it might participate in squid cartilage proteoglycan aggregate formation.

Occurrence of a novel collagen with three distinct chains in the cranial cartilage of the squid Sepia officinalis: comparison with shark cartilage collagen

A unique collagen with three distinct chains, was purified from the cranial cartilage of the squid Sepia officinalis, by pepsinisation and salt precipitation and compared with shark cartilage collagen. These chains, which were different from the known cartilage collagen chains, were referred as C1, C2 and C3, had approximate molecular weights of 105 kDa, 115 kDa and 130 kDa, respectively, and were present in a ratio of 3:2:1, suggestive of two molecules of composition, [(C1) 2C2] and [C1C2C3]. These collagens were purified by fractionation at acid and neutral pH, and by ammonium sulfate precipitation. Solubility data indicated that this collagen was more crosslinked than the type I collagen isolated from cartilage of shark, Carcharius acutus. In vitro fibrillogenesis revealed that the sepia collagen formed denser aggregates, as compared to shark collagen, and was stabilised by a higher degree of carbohydrate association. Polyclonal antisera raised against shark collagen was also reactive against the sepia collagens, while the converse was not true, indicating the high immunospecificity of the latter. These results demonstrate collagen polymorphism in an invertebrate cartilage and may hold significance in understanding tissue calcification and molecular evolution. Further, these collagens may represent ancestral forms of vertebrate minor collagens like typeV/XI.

Cranio-facial skeletal development in three human synophthalmic holoprosencephalic fetuses

In three human fetuses with synophthalmic holoprosencephaly (8, 14, 23 wks. p.c.) and two normal human fetuses (9 and 13 wks. p.c.) the anatomy of the cranial base, facial cranium and their relation to the notochord was studied using serial histological sections and computer aided three-dimensional reconstruction methods. Mesethmoidal cartilage differentiation was variably deficient in all three holoprosencephalic cases. The premaxillary bones were rudimentary with missing tooth buds. The development of the sphenoid bone was defective in two of the holoprosencephalic cases (8, 14 wks. p.c.). The notochord terminated normally within the sphenoid body in all investigated cases. Our results indicate that in holoprosencephaly there is a general defect in the midline cranial cartilage differentiation rostral to the notochord.

Localization of IGF-I and IGF-I Receptor mRNA inSparus aurataLarvae

Studies of the ontogeny of IGF-I mRNA during embryonic and larval development of the gilthead sea breamSparus auratashowed its expression during these early developmental stages. The present study appliesin situhybridization to localize IGF-I and IGF receptor mRNAs in 16-day larvae ofS. aurata.Paraffin sections were hybridized with homologous RNA probes labeled by [35S]UTP. IGF-I mRNA expression was found mainly in chondrocytes, in both the gill arches and cranial cartilage, in skeletal muscle, in the brain, in the pancreas, in the retina, and in the epithelial cells surrounding the lens. A strong positive reaction for IGF receptor mRNA was found in skeletal muscle, in the pancreas, and in the lymphoid tissue found in the intertubular tissue of the kidney. Signals were less intense in brain and chondrocytes. It is suggested that in teleosts, as in higher vertebrates, IGF-I may be involved in the regulation of tissue growth and differentiation in an autocrine/paracrine manner.

Chapter 3 Manipulations of Neural Crest Cells or Their Migratory Pathways

Publisher Summary This chapter discusses the manipulations of neural crest cells or their migratory pathways. The formation of the embryo involves intricate cell movements, cell proliferation, and differentiation. The neural crest has long served as a model for the study of these processes, because neural crest cells undergo extensive migrations and give rise to many diverse derivatives. Neural crest cells arise from the dorsal portion of the neural tube. Several unique properties of these cells make the neural crest an ideal system for studying cell migration and differentiation. First, these cells migrate extensively along characteristic pathways. Second, they give rise to diverse and numerous derivatives, ranging from pigment cells and cranial cartilage to adrenal chromaffin cells and the ganglia of the peripheral nervous system. In addition, their characteristic position of premigratory neural crest cells within the dorsal portion of the neural tube makes them accessible to surgical and molecular manipulations during the initial stages in their development.

ULTRASONOGRAPHIC ANATOMY OF THE NORMAL CANINE STIFLE

Ultrasonographic examination of the normal canine stifle joint was performed to characterize its normal anatomy. Stifles of four normal adult dogs were imaged in sagittal and transverse planes and each anatomic structure visualized was recorded. Normal anatomic structures consistently seen included the patellar tendon, medial and lateral menisci, the cranial cruciate ligament and femoral condyle cartilage. The caudal cruciate ligament was visualized in two dogs. Collateral ligaments and meniscal ligaments were not visualized. The dogs were then euthanized and each stifle was isolated. Following removal of superficial muscles and skin, each stifle was imaged in a water bath to definitively identify the structures that had previously been visualized on the live dogs. The ultrasonographic appearance of the isolated stifle specimens was similar to that found in live dogs. The results of this study indicate that ultrasound can be used to image the normal anatomy of the canine stifle. The echogenicity of the patellar ligament, cruciate ligaments, menisci and articular cartilage was similar to that previously reported in equine stifles and human knees.

Growth changes in the craniofacial complex of the rat after prolonged papain administration

The purpose of the study was to evaluate by radiographic and biometric means the effects of prolonged papain administration on growth of the craniofacial complex, to gain further insight into the importance of cranial cartilage for skull enlargement during rapid skeletal growth (25 to 70 days of age). Fifty 21-day-old male Lewis strain rats were divided randomly into three groups. Group 1 ( n = 20) received 45 daily injections intraperitoneally of a 2% solution of crude papain in normal saline from 25 to 70 days of age. Group 2 ( n = 20) were given 30 daily injections at the same dose level from 40 to 70 days of age, and group 3 ( n = 10) served as untreated controls. Animals were weighed daily until 70 days of age and then every other day until killed at day 120. Submental-vertex and lateral skull radiographs were obtained at 25, 40, 53, 70, 90, and 120 days of age with standardized skull positioning and radiographic settings. Cephalometric measurements of 22 linear dimensions were made in each of 360 cephalograms, and the data analyzed statistically with Student's t test. Growth velocity curves were evaluated for evidence of catch-up growth. Body-weight gain by papain-treated groups was less than for controls (p < 0.001). Skull and nasal lengths, middle cranial base length, and sphenoidal length were significantly shorter (p < 0.001). Neurocranial length was also shortened, whereas neurocranial height and supraoccipital height increased. Reduction of skull dimensions was generally more pronounced in the earlier injected group. The effectiveness of catch-up growth was not constant and was more successful in the later injected group. Catch-up growth was effective for posterior nasal, neurocranial and supraoccipital heights, cranial width, and posterior cranial base length. The incomplete nature of catch-up growth may have been due to the particular timing and severity of the insult, the frequency of administration of the enzyme and the peak velocity of dimensional growth relative to the time of papain administration. (A M J O RTHOD D ENTOFAC O RTHOP 1994;105:270-8.)

Self-aggregation of squid cranial cartilage proteoglycans

Squid cranial cartilage has been found to contain three different proteoglycan populations, two of which form aggregates (Vynios, D. H., Tsiganos, C. P., Biochim Biophys Acta 1033: 139–147, 1990). The aggregation involves interaction of their protein cores as assessed by electron microscopy and biochemical data. Aggregating oligopeptides were isolated after mild trypsin digestion which inhibited self-aggregation of proteoglycans. The aggregation does not involve interaction of the side chains of polar amino acids and evidence is provided that it is mediated through hydrophobic interaction. It is enhanced upon concentration or incubation of the samples at 37°C.

Three Myxosporeans Found in the Cranial and Branchial Tissues of Rainbow Trout in California

Abstract Three myxosporeans were encountered in the cranial tissues of a California population of rainbow trout Oncorhynchus mykiss examined for the presence of Myxobolus cerebralis, the causative agent of whirling disease. Typical spores of M. cerebralis and a previously undescribed species of Myxobolus were found in the cranial tissues prepared by the pepsin HCl-trypsin digestion method. Henneguya zschokkei was also detected in digest preparations of cranial tissues, but was more numerous when branchial cartilage was included in the preparations. Microscopic examinations of tissues of individual rainbow trout showed occasional infections with both myxobolid species. Myxobolus cerebralis trophozoites and spores were found in the cranial and gill cartilage, and Myxobolus sp. was found in the brain and spinal cord. Henneguya zschokkei was also found within granulomas in the connective tissues below the gill arch. Both M. cerebralis and H. zschokkei were associated with a chronic inflammatory response in th...

Sensory evoked potentials in unanesthetized unrestrained cuttlefish: a new preparation for brain physiology in cephalopods

Up to five microelectrodes inserted through short hypodermic needles in the cranial cartilage of Sepia officinalis recorded potentials while the cuttlefish moved freely in a small enclosure. Compound field potentials and unit spikes were seen during ongoing, spontaneous activity and after sensory stimulation. Ongoing activity resembles that reported for octopus, with maximum power usually below 20 Hz. Amplitude varies greatly but has not been seen to shut off or turn on abruptly and globally as in octopus. Evoked potentials, focally large after flashes of light consist of several waves; the first is largest, positive and peaks at ca. 35 ms (called P35), followed by ca. P75, P95, N110 and smaller waves or oscillations lasting more than 0.5 s. The Upper Following Frequency (highest flashing rate the potentials can follow 1:1), without averaging, is greater than 15 flashes/s (20-22 degrees C); at 20/s the 1:1 following lasts for 1 or 2 s. The Lower Fusion Frequency of averaged responses is less than 30/s. Gentle tapping of the tank wall evokes local, brief, fast potentials. No responses have been found to loud air-borne clicks and tone bursts with principal energy at 300 Hz or to electric fields in the bath at 50-100 microV/cm. In a few loci relatively large slow Omitted Stimulus Potentials have been seen following the end of a train of flashes at more than 5/s; these are by definition event related potentials and a special, central form of OFF response.

Squid proteoglycans: isolation and characterization of three populations from cranial cartilage

Squid cranial cartilage is poor in proteoglycans. They were extracted by 2% SDS and purified by isopycnic centrifugation in the presence of detergent. According to their buoyant density and hydrodynamic size they were fractioned inte three structurally different populations of M r 1.3 · 10 6, 0.6 · 10 6 and 1.0 · 10 6. The proteoglycans of each population differ in the number of oversulphated chondroitin sulphate chains, ranging from two to five, in the number and size of uronic acid and sulphate containing oligosaccharides and in the size of their core protein. The majority, if not all, of the oligosaccharides are linked to the protein via an O-glycosidic bond involving galactosamine and most likely xylose. The chondroitin sulphate chains are segregated on a small peptide segment of the molecule which also contains a large proportion of the oligosaccharides. The proteoglycans have no tendency to interact with hyaluronate.

Morphological integration in the cranium during anuran metamorphosis.

We examined the role of thyroid hormone in mediating morphological integration between cranial cartilage and bone during anuran metamorphosis. Exogenous T3 applied to premetamorphic tadpoles (Bombina orientalis) via intracranial implants of plastic micropellets precociously induced typical metamorphic changes in both tissues, but also dissociated the relative timing of developmental events between them. Morphological integration between the two primary cranial tissues is achieved in part by each tissue responding independently to endocrine factors and does not reflect a tight developmental coupling between them.

Squid cartilage collagen: Isolation of type I collagen rich in carbohydrate

1. 1. The cranial cartilage of the squid Todarodes pacificus, which resembles vertebrate hyaline cartilage in its microscopic appearance, was solubilized by limited proteolysis with pepsin. 2. 2. Characterization of the solubilized collagen suggests that the squid cartilage contains primarily Type I collagen which is rich in hydroxylysine-linked carbohydrate, similar to the cartilage-type collagen of vertebrates. 3. 3. Tissue-specific differences in proline hydroxylation and thermal stability of squid Type I collagen were observed, comparing the collagens of cranial cartilage and mantle skin tissues.

Proteoglycans from squid cranial cartilage: extraction and characterization

Abstract 1. 1. The majority of proteoglycans (85% of uronic acid) were extracted from squid cranial cartilage with 2% SDS. 2. 2. Conventional methods with 4 M Gdn HCl alone or with 4% Zwittergent, extracted only 30%. 3. 3. The proteoglycans were isolated by density gradient centrifugation. 4. 4. At least 25% of the extracted proteoglycans with 2% SDS do not form aggregates with hyaluronate. 5. 5. The proteoglycan monomers are of varying hydrodynamic size, those extracted with 4 M Gdn HCl being the smallest of average molecular weight 750,000.

Analysis of polysulfated chondroitin disaccharides by high-performance liquid chromatography

A high-performance liquid chromatography method for analyzing disaccharides derived from chondroitin sulfate glycosaminoglycans has been developed which employs a Whatman Partisil-10 PAC amino-cyano column and an acetonitrile/methanol/ammonium acetate solvent to resolve disulfated, monosulfated, and unsulfated disaccharides in a chromatographic run of less than 20 min. The single known trisulfated chrondroitin disaccharide can be eluted in an alternate solvent system containing the same mobile phase components in different proportions. Disaccharides were prepared for chromatography from glycosaminoglycans and proteoglycans of known compositions by digestion with chondroitinase ABC, with the exception of king crab cartilage glycosaminoglycan which was incubated sequentially with hyaluronidase and chondroitinase ABC. Disaccharides were extracted from the digestion mixtures in 80% ethanol, dried over nitrogen, resuspended in HPLC solvent, and chromatographed at a flow rate of 1 ml/min. Unsaturated disaccharides in the column eluate were detected by continuous ultraviolet absorbance monitoring at 232 nm; alternatively, fractions were collected and assayed for uronic acid content or radioactivity. By utilizing the HPLC technique in conjunction with chondroitinase ABC and AC digestion and sulfatase hydrolysis, the epimeric structures of chondroitin sulfates E and H were confirmed. With this technique, rapid and reproducible analyses of chondroitin sulfate disaccharides generated from mouse mast cell proteoglycan and from glycosaminoglycans of squid cranial cartilage, shark skin, hagfish skin, and hagfish notocord were in close agreement with compositions obtained by other techniques.

Analysis of the acid polysaccharides from squid cranial cartilage and examination of a novel polysaccharide

The polysaccharides of cranical cartilge were isolated by ethanol precipitation after papain digestion and β-elimination procedures and were fractionated chromatographically on CPC-cellulose. In addition to the previously described, heavily oversulphated chondroitin sulphate, the tissue contained small amounts of hyaluronic acid, which, however, co-eluted with the chondroitin sulphate from the CPC-cellulose. Approx. 20% of the isolated polysaccharides consisted of an acidic polysaccharide which to our knowledge is not previously described. This polysaccharide consists mainly of glucuronic acid, galactose and mannose in a molar ratio of 1:2:1. Gel chromatography of the preparation indicated a polydisperse molecule with an apparent average molecular weight of 39200 on weight basis ( M w ) and 31400 on number basis ( M n ) .

Differential changes in cartilage cell proliferation and cell density in the rat craniofacial complex during secondary palate development.

During mammalian secondary palate formation sagittal growth of the lower face has been shown to be more rapid than that of the upper face, and the tongue and mandible extend beneath the primary palate. In order to identify factors contributing to this differential growth pattern, cellular and morphologic growth of the major cartilages of the upper and lower facial regions were studied in radioautographic sections labeled with tritiated thymidine. Evaluation of cell-density recordings, labeling indices, and structural dimensions revealed significant differences between Meckel's cartilage in the lower face, and the nasal cartilage and anterior cranial base cartilage in the upper face. After formation of the precartilaginous blastema, labeling indices were high in Meckel's cartilage (20-30%), but very low in the nasal cartilage and the anterior cranial base (0-2%). During secondary palate formation the volume of Meckel's cartilage increased more rapidly than the other cartilages and its growth was primarily in the sagittal direction. Between days 15 and 17, the increase in the length of Meckel's cartilage (165%) was approximately twice as great as the increase in the combined length of the nasal cartilage and the anterior cranial base (77%). During this period induction of cleft palate with some teratogens has been shown to severely retard growth of Meckel's cartilage and produce mandibular retrognathia that contributes to delayed elevation of the palatal shelves. Therefore, extensive cell proliferation in Meckel's cartilage, during a period of limited proliferation in other craniofacial cartilages, appears to contribute to its rapid growth and its differential sensitivity to growth inhibition.

In vitro accumulation of mineral components by invertebrate cartilage.

In vitro mineralization of the gill cartilage of Limulus (horse-shoe crab) has been reported previously (Eilberg et al. 1975). The present study demonstrates that cranial cartilage of Loligo (squid) and odontophore cartilage of Busycon (marine snail) also mineralize in vitro in hydroxyapatite-metastable media, but not in carbonate-metastable media. In all three of these cartilages, light phase-contrast microscopy revealed that the mineral phase occured in the form of spherical or ovoid granules ranging between 2 and 9 mum in diameter. During mineralization, the granules appeared successively in the perichondrium, in the matrix, and finally, within chondrocytes. Mineralization occurred more rapidly in Eusycon odontophore cartilage which has a significantly higher content of phosphatidyl serine than in Limulus gill or Loligo head cartilages. In all tissues the mineralization process is related to temperature, taking place most actively at 37 degrees C and only weakly at 50 degrees C.

Lipid components and in vitro mineralization of some invertebrate cartilages.

Cartilage from several invertebrate organisms Limulus polyphemus (Arthropoda), Eudistylia polymorphia (Annelida), Busycon canaliculatum (Mollusca) and Loligo pealii (Mollusca) were assayed for RNA, DNA, collagen, noncollagenous protein and lipid content. The variability in levels of collagen (1-19%) in the respective tissues, appeared to correlate well with their relative matrix content as revealed by previous histologic study. Determinations were made of mono- and diglycerides, triglycerides, free fatty acids, methyl esters, steroid-like materials and steroid-like esters, phosphatidyl choline, phosphatidyl serine, phosphatidyl ethanolamine, lysolecithin, phosphatidic acid, and sphingomyelin. The fatty acid composition of the major sub-groups was determined, and it was found that the ordinary fatty acids, palmitic, stearic, and oleic represented the largest fraction, whereas values for unsaturated fatty acids in the invertebrate cartilages were smaller than those usually reported for marine organisms. Significant levels of phosphatidyl serine were found in all tissues, being especially high in the odontophore cartilage of Busycon, i.e., 13.7% of phospholipid content. Some unusual and as yet unidentified fatty acids, representing approximately 10% of the total fatty acid composition were also found. In vitro mineralization of Busycon odontophore cartilage, Limulus gill cartilage and Loligo cranial cartilage were carried out by incubation of the respective tissues at 37° C in a medium metastable for hydroxyapatite. Of the three tissues mentioned above, earliest signs, and most rapid mineralization occurred in the Busycon cartilage, which also has the highest content of phosphatidyl serine. This latter observation is of interest because of the demonstrated role of phosphatidyl serine in mineralization of vertebrate cartilages, and suggests the possibility of a similar role in the in vitro invertebrate cartilage mineralization.

Note on a new host of Myxobolus aeglefini

Some time ago Mr E. Wilson, of the Marine Laboratory, Aberdeen, brought to notice the abnormal appearance of the cranial cartilage in a plaice (Pleuronectes platessa L.) which had been tagged (in St Andrews Bay) and subsequently recaptured (24 miles south-south-east of Arbroath). The fish was a 44·5 cm. long female, 6 years old, with mature ovary and in good condition generally. The cartilage, exposed by the opening of the cranium to remove the otoliths, seemed hypertrophied and full of whitish splotches giving it a mottled appearance. These ‘splotches’ were formed by irregular, vein-like branching canals, in many places along their length swelling into cyst-like, subspherical cavities up to 3 mm. in diameter. They were filled by a mass of soft substance which was found to be an accumulation of Cnidosporidian spores. Small granules of the substance were also found inside the otic capsules. It is possible, however, that these granules were displaced into the capsules during the opening of the cranium.