The study of bone microstructure of fossil vertebrates (i.e., paleohistology) has demonstrated to be a very important source of paleobiological information. Paleohistological studies are based on the standardized analysis of petrographic thin sections. Although the development of new technologies (e.g., microtomography) have provided non-destructive procedures for the study the fossil tissues, thin sections are still the main source of information in paleohistology. In this contribution, we provide a detailed protocol for sampling and thin-sectioning preparation of bone tissue from both fossil and extant vertebrates. We describe the most common procedures for sampling and also some particularities related to variations in equipment and sampling techniques. The main goal of this contribution is to offer an alternative protocol for research teams of recent formation and/or with limited funding.
Sharpey's fibers are considered the anatomical structures integrated to the muscles. Since these fibers leave marks at the microscopic level, their presence and distribution are used as evidence of muscle attachment in extinct and extant forms. In recent years, studies have been focusing on muscle–bone and tendon–bone interaction mostly on mammals. The main objective of this work is to contribute to the morphological and histological knowledge of muscle attachment in other amniotes, such as reptiles, and their variation related to different locomotor habits. In this way, a study was performed on terrestrial and aquatic turtles. The musculature related to the movement of the humerus, and pectoral girdle in Chelonoidis chilensis, Phrynops hilarii and Hydromedusa tectifera was analyzed. Dissections were performed mapping the origins and insertions of each muscle and undecalcified thin sections were performed in specific muscular attachment sites. We found some differences which were not previously reported, related to the insertion of the m. pectoralis, the m. coracobrachialis magnus and the origin of the m. tractor radii. The osteohistology revealed the presence of Sharpey's fibers in the cortex of all the bone elements analyzed. Patterns were established in relation to the orientation and density of Sharpey's fibers, which were used for the categorization of each muscle attachment site. The comparative micro‐anatomical study of these areas did not reveal any important differences between terrestrial and freshwater turtles in muscles involved with the rotation, abduction and adduction of the humerus. In this way, the preliminary results suggest an absence of correlation between the distribution and density of Sharpey's fibers between different habitat forms, at least in the bones and species analyzed.
This article presents a detailed comparative analysis of the bone microstructure of three extant species of South American turtles. The main histological characteristics of postcranial bones are identified, as well as the intraskeletal, ontogenetic and interspecific variation between aquatic and terrestrial species. For this purpose, thin sections of postcranial bones (seventh cervical vertebra, coracoid, scapula, humerus, radius, ulna, ischium, ilium, pubis, femur, tibia, and fibula) of juvenile and adult specimens of aquatic (Phrynops hilarii and Hydromedusa tectifera) and terrestrial (Chelonoidis chilensis) turtles were analyzed. Bone histology revealed an intraskeletal variation of the microanatomical and microstructural organization in these turtles. The cortical bone is composed of poorly vascularized lamellar and parallel‐fibered bone tissue interrupted with lines of arrested growth (LAGs), reflecting a cyclical slow growth rate throughout these turtles' life. Although in the adult specimens a growth rate decrease was observed, none of them have reached somatic maturity. The juvenile and the adult of Chelonoidis chilensis, unlike the aquatic species studied, presented a higher vascularization in their bones, which could imply a faster growth rate in this land specie. The number of LAGs was higher in the stylopodial and zeugopodial bones, which would make these elements suitable for approximate age estimations. Pectoral and pelvic girdle bones also exhibited a good record of LAGs. The information here obtained on extant species represents a powerful tool for the interpretation of paleobiological traits present in closely related fossil forms.
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