Consequences of ankle joint immobilisation: insights from a morphometric analysis about fibre typification, intramuscular connective tissue, and muscle spindle in rats.

Orthosis immobilisations are routinely used in orthopaedic procedures. This intervention is applicable in bone fractures, ligament injuries, and tendonitis, among other disorders of the musculoskeletal system. We aimed to evaluate the effects of ankle joint functional immobilisation on muscle fibre morphology, connective tissue, muscle spindle and fibre typification triggered by a novel metallic orthosis. We developed a rodent-proof experimental orthosis able to hold the tibiotalar joint in a functional position for short and long terms. The tibialis anterior muscles of free and immobilised legs were collected and stained by histology and histochemistry techniques to investigate general muscle morphology, connective tissue and muscle fibre typification. Morphometric analysis of muscle cross-section area, fibre type cross-section area, fibre type density, percentage of intramuscular connective tissue, and thickness of the muscle spindle capsule were obtained to gain insights into the experimental protocol. We found that short- and long-term immobilisation decreased the cross-section area of the muscles and induced centralisation of myonuclei. The connective tissue of immobilised muscle increased after 2 and 4 weeks mainly by deposition of type III and type I collagen fibres in the perimysium and endomysium, respectively, in addition to muscle spindle capsule thickening. Type IIB muscle fibre was severely affected in our study; the profile assumed odd shapes, and our data suggest interconversion of these fibre types within long-term immobilisation. In conclusion, our protocol has produced structural and histochemical changes in muscle biology. This method might be applied to various rodent models that enable genetic manipulation for the investigation of muscle degeneration/regeneration processes.

Influence of rainfall and temperature on the spermatogenesis of Leptodactylus macrosternum (Anura: Leptodactylidae)

In the semi-arid environment, the reproductive success of anurans depends on adaptations in their life cycle, which synchronizes with ideal environmental conditions to maximize the number of offspring. In this study changes in the histological and morphometric aspects of the testes of Leptodactylus macrosternum Miranda-Ribeiro, 1926 are characterized, to evaluate the influence of rainfall and temperature on them. Specimens were collected at Horto Florestal Olho dÁgua da Bica - HFOB (06°4920”S, 36°1585”W) area, municipality of Cuité, state of Paraíba, Brazil. Search for specimens was active, happened at night, and amounted to 15 days from January to December 2013. The densities of spermatids (primary and secondary), sperm and area of the seminiferous locules of the testes were registered. The influence of climate variables (rainfall and temperature) on the density of primary and secondary spermatids, sperm and locular area were verified using Simple Linear Regression. Primary spermatids had the lowest density in July (57.90 ± 51.54 mm2), with a peak in November (300.32 ± 117.35 mm2); secondary spermatids had the lowest density in December (287.87 ± 79.05 mm2), with a peak in May (135,727.00 ± 301.13 mm2); sperm was in the lowest density in July (237.37 ± 121.10 mm2), with a peak in June (2,270.45 ± 602.62 mm2) and the locular area had the lowest density in December (40,292.9 ± 8,174.20 µm2) and highest density in June (338,875.01 ± 2,262.10 µm2). A notable decrease in sperm density was evident between June and July. That, associated with the observation of a larger locular area in June, allowed us to identify as June as the peak of spermatogenesis and the following month as the most potentially reproductive. The density of secondary spermatids(r = 0.02), sperm (r = 0.21) and locular area (r = 0.01) showed dependency on rainfall whereas only sperm (r = 0.09) showed dependency on temperature.[...](AU)

Influence of rainfall and temperature on the spermatogenesis of Leptodactylus macrosternum (Anura: Leptodactylidae)

In the semi-arid environment, the reproductive success of anurans depends on adaptations in their life cycle, which synchronizes with ideal environmental conditions to maximize the number of offspring. In this study changes in the histological and morphometric aspects of the testes of Leptodactylus macrosternum Miranda-Ribeiro, 1926 are characterized, to evaluate the influence of rainfall and temperature on them. Specimens were collected at Horto Florestal Olho d’Água da Bica - HFOB (06°49’20”S, 36°15’85”W) area, municipality of Cuité, state of Paraíba, Brazil. Search for specimens was active, happened at night, and amounted to 15 days from January to December 2013. The densities of spermatids (primary and secondary), sperm and area of the seminiferous locules of the testes were registered. The influence of climate variables (rainfall and temperature) on the density of primary and secondary spermatids, sperm and locular area were verified using Simple Linear Regression. Primary spermatids had the lowest density in July (57.90 ± 51.54 mm2), with a peak in November (300.32 ± 117.35 mm2); secondary spermatids had the lowest density in December (287.87 ± 79.05 mm2), with a peak in May (135,727.00 ± 301.13 mm2); sperm was in the lowest density in July (237.37 ± 121.10 mm2), with a peak in June (2,270.45 ± 602.62 mm2) and the locular area had the lowest density in December (40,292.9 ± 8,174.20 µm2) and highest density in June (338,875.01 ± 2,262.10 µm2). A notable decrease in sperm density was evident between June and July. That, associated with the observation of a larger locular area in June, allowed us to identify as June as the peak of spermatogenesis and the following month as the most potentially reproductive. The density of secondary spermatids(r = 0.02), sperm (r = 0.21) and locular area (r = 0.01) showed dependency on rainfall whereas only sperm (r = 0.09) showed dependency on temperature.[...]