The descriptive anatomy of the cricoarytenoid articulation provides an essential foundation for understanding disorders of mobility of the larynx, especially in carcinology. Thirteen formaline-preserved anatomic specimens of the adult larynx were studied and 4 pathologic larynges with loss of mobility due to a malignant tumor. The cricoid and arytenoid articular surfaces showed major intra- and inter-individual variations, causing dynamic asymmetry at the glottic level. They were joined by a connective-elastic articular capsule bounding a cavity, characterized by a pseudo-meniscal synovial ridge and deep peripheral blind recesses, indicative of great articular mobility. The cricoarytenoid ligament shares in stabilizing the articulation. The posterior cricoarytenoid m. (abductor) and the lateral cricoarytenoid m. (adductor) have a motor innervation derived from the inferior laryngeal nerve, which forms an endolaryngeal arch with a ventral concavity, in contact with the lateral articular recess. The cricoarytenoid articulation thus appears as a diarthrosis possessing three degrees of liberty during movements of glottic abduction and adduction: an antero-posterior rocking movement, an antero-medial shift of the arytenoid on the cricoid, and a less marked axial rotation. Histological study of the cricoarytenoid articulation where mobility was reduced by carcinomatous infiltration showed that each articular component may be affected (muscles, cartilage, capsule, nerve), and that several components may be involved simultaneously to a minimal degree. The therapeutic implications are important, particularly in conservative laryngeal surgery.
The viscous seminal plasma (SP) is currently a major impediment to the handling of ejaculate and the development of some biotechnologies in South American camelids. The vas deferens-collected spermatozoa of alpacas is a useful technique to avoid this problem. On the other hand, SP contains a large protein component that has been implicated in the function of spermatozoa within the female reproductive tract. In this sense, the low fertility achieved using transcervical insemination with frozen-thawed spermatozoa in alpacas could be improved by adding SP. This study aimed to evaluate the effect of the whole SP on some in vitro parameters of alpaca spermatozoa after the freezing-thawing-process and the fertility after artificial insemination. It would contribute to a better understanding of the interaction between thawed sperm cells and SP. Spermatozoa were obtained by surgically diverted vas deferens. The samples were diluted with a Tris-based extender, packaged in straws, and frozen. At thawing, each straw was divided into two post-thawing conditions: with the addition of 10% of PBS (control) or with 10% SP (treatment). The sperm cells were evaluated using dynamic parameters, sperm cell morphology, and morphometry. Fertility was assessed by an artificial insemination trial. All in vitro parameters were analyzed by ANOVA. A heterogeneity test was scheduled for the fertility trial. After the freezing-thawing process, motility and plasma membrane functionality was improved when SP was added. No differences were found for post-thaw viability between the control and treatment samples. The percentage of normal cells was higher with SP at post-thawing, and a decrease of the presence of bent tailed spermatozoa with a droplet in the SP group was observed. The length of the head spermatozoa was 3.4% higher in the samples with PBS compared to those in which SP was added. Females pregnant at day 25 post-insemination were 0/12 (with SP inside the straw) and 1/10 (without SP inside the straw). In conclusion, the presence of 10% SP at post-thawing improves sperm cells' motility, functionality, and morphology, indicating that it would be beneficial to improve the frozen-thawed alpaca's physiology spermatozoa. More fertility trials must be developed to increase this knowledge.
KEY WORDS:Viabilidad in vitro e in vivo de los espermatozoides congelados/descongelados del conducto deferente de alpacas (Vicugna pacos)Viability in vitro and invivo of sperm frozen/thawed from alpaca (Vicugna pacos) vas deferens El objetivo del presente estudio fue evaluar la viabilidad in-vitro e in-vivo de los espermatozoides procedente de los conductos deferentes de alpacas. Se utilizaron 2 reproductores como donadores de espermatozoides con desviación del conducto deferente. Los espermatozoides colectados se sometieron a la congelación y descongelación con el dilutor Triladyl®. Durante el procesamiento de los espermatozoides se evaluaron la motilidad y la prueba hipo-osmotica. En la inseminación se evaluó la proporción de gestaciones producidas. Los resultados fueron los siguientes: La motilidad de los espermatozoides para los reproductores 1 y 2 fueron: a los 37°C 65.16 y 63.37% sin diferencia (P>0.05), al enfriamiento (5°C) 52.63 y 48.31% similares (P>0.05) ya la descongelación 24.51 y 33.18% mostraron diferencia (P<0.05). A la prueba hipo-osmótica fueron; a los 37°C 51.55 y 52.98% similares (P>0.05), Al enfriamiento (5°C) 46.67 y 55.93% mostraron diferencia (P<0.05) y a la descongelación 20.06 y 32.18% con diferencia (P<0.05). Las gestaciones fueron: Con espermatozoides frescos diluidos 36.36% (4/11), con espermatozoides descongelados 25.00% (5/20) y con monta 54.54% (6/11), siendo similares (P>0.05). En conclusión los espermatozoides procedentes de los conductos deferentes soportan el congelamiento y a la inseminación artificial producen gestaciones.The aim of this study was to evaluate the in-vitro and in-vivo viability of sperm from the vas deferens alpacas. 2 males as sperm donors were used to offset the vas deferens. The collected sperm cells were subjected to freezing and thawing with dilutor Triladyl®. During processing of sperm motility and hypo-osmotic test they were evaluated. Insemination in the proportion of pregnancies produced was evaluated. The results were as follows: The sperm motility of males 1 and 2 were: at 37 ° C 65.16 and 63.37% with no difference (P> 0.05), cooling (5 ° C) 52.63 and 48.31% similar (P > 0.05) and thawing 24.51 and 33.18% showed no difference (P <0.05). The hypo-osmotic test were; at 37 ° C 51.55 and 52.98 similar percentage (P> 0.05), and cooling 55.93% 46.67 showed no difference (P <0.05) and thawing 20.06 and 32.18% with difference (P <0.05). Pregnancies were diluted with fresh sperm 36.36% (4/11), with defrosted sperm 25.00% (5/20) and 54.54% mounts (6/11), being similar (P> 0.05). In conclusion sperm from the vas deferens withstand freezing and insemination are able to produce pregnancies.
El objetivo del estudio fue determinar el efecto de tres curvas de congelación sobre la viabilidad pos-descongelación de espermatozoides colectados del conducto deferente de llamas. Se utilizaron seis llamas machos con desviación quirúrgica de los conductos deferentes. Las muestras de los seis machos se mezclaron para su procesamiento (pools) y se diluyeron con Tris-yema de huevo. Se procedió al enfriamiento hasta los 5 °C donde se completó la dilución (dilutor con glicerol) y se mantuvo por media hora (fase de equilibrio). Las muestras en pajillas de 0.25 ml fueron sometidas a congelamiento utilizando una tasa de descenso de temperatura de -20°C/min hasta llegar a -80°C (TI), a -100 °C (TII) y a -120 °C (TIII) en 4, 5 y 6 minutos, respectivamente, para finalmente almacenarlas en nitrógeno líquido. Las muestras fueron colectadas durante tres meses (n=19 pooles). Se determinó la motilidad total (motilidad progresiva, circular, oscilatoria), viabilidad y funcionalidad de membrana luego de la colecta, en la fase de equilibrio y al descongelamiento. Se observó una disminución significativa (p˂0.05) en todas las características espermáticas evaluadas en las muestras equilibradas respecto a las muestras luego de la colecta. Se obtuvo una correlación alta positiva entre viabilidad y motilidad total (r2=0.78) y en fase de equilibrio entre funcionalidad de membrana y motilidad total (r2=0.881) En las muestras descongeladas, la motilidad total y viabilidad fueron significativamente mayores en las muestras congeladas con la curva de congelamiento TIII respecto al TI (p=0.041 y p=0.003, respectivamente). No se observaron diferencias significativas en la funcionalidad de membrana entre las tres curvas de congelamiento (p˃0.27). En conclusión, la curva de descenso de la temperatura hasta los -120 °C utilizando una tasa de -20 °C/min sería la más adecuada para criopreservar espermatozoides de llama obtenidos a partir de la desviación de los conductos deferentes.
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