Parameters of the Reproductive Tract, Spermatogenesis, Daily Sperm Production and Major Seminal Plasma Proteins of Tropically Adapted Morada Nova Rams

Abstract

This study describes the reproductive parameters of Morada Nova rams, a breed of hair sheep from Brazil and with unique adaption to tropical environments. At 42 weeks of age, 15 rams were subjected to semen collection and, 1 week later, animals were slaughtered for collection of testes, epididymis and accessory sex glands. We conducted 2-D electrophoresis of seminal plasma proteins and major spots of stained gels were identified by LC-MS/MS. Total RNA was isolated from testis, epididymis and vesicular glands and subjected to qPCR. At slaughter, scrotal circumference and testicular weight were 27.5 ± 0.5 cm and 109.5 ± 6.0 g, respectively. Seminiferous tubule (ST) diameter was 188.3 ± 4.0 μm and each testis contained 1.9 ± 0.1 Sertoli cells (×10(9) ). Each Sertoli cell supported 0.1 ± 0.01 A spermatogonia, 3.0 ± 0.2 pachytene spermatocytes and 7.7 ± 0.5 round spermatids/tubule cross section. Daily sperm production reached 5.6 × 10(6) cells/g of testis parenchyma. Testis size appeared as indicative of ST diameter and associated with epididymal measurements, as well as with the population of round spermatids and Sertoli cells/testis. Rams with heavier testes had greater daily sperm production and more Sertoli cells/testis. We detected 90.9 ± 9.6 spots per 2-D gel of seminal plasma. Major seminal proteins were identified as ram seminal vesicle proteins at 14 and 22 kDa, representing 16.2% and 12.8% of the total intensity of valid spots in the gels, respectively. Expression of both genes was greater in the vesicular glands as compared to testis and epididymis. Pixel intensity for those proteins in the 2-D gels was significantly correlated with seminal vesicle weight. This is the first description of the basic reproductive aspects of Morada Nova rams, including protein profiles of their seminal plasma. These findings will allow a better understanding of their reproductive physiology.

Testicular morphometry as a tool to evaluate the efficiency of immunocastration in lambs

“…The S/PQ ratio obtained in the control group corroborates the findings of Sousa et al (2014) in Morada Nova sheep, whereas the S/Ar ratio was higher than that reported by the authors. The total production of PQ is usually adjusted numerically to support the capacity of Sertoli cells, causing the degeneration of large numbers of cells ( De Rooij and Lok, 1987 ).…”

Annual variation in reproductive parameters and sexual behaviour of Saint Croix rams in a semi-desert region in Mexico

“…The T levels observed in this study are similar to the values reported in Saint Croix rams in Mexico (Aguirre et al 2007) and Morada Nova rams in Brazil (Sousa et al 2014). However, when raised in tropical regions, the effect of season on T is more nutritionally dependent, especially in the case of cross-breeding with wool breeds (Zhang et al 2005;Cárdenas-Gallegos et al 2015).…”

Gene and protein expression in the reproductive tract of Brazilian Somalis rams

“…After thawing, samples were macerated, homogenized and incubated for 2 hr under moderate agitation with a lysis buffer containing 20 ml PBS (1x), 200 μl Triton X100, 20μL of the 500μL protease inhibitor (Sigma Aldrich, EUA). Subsequently, the material was centrifuged (4°C, 10,000 g) and the supernatant, transferred to new tubes (Souza et al, ). Then acetone (1:9) was added, centrifuged (5 min., 1,000 g, 4°C) and the pellet was solubilized in 0.5 ml PBS (1x).…”

“…The mRNA expression level evaluated in technical replicates weresubjected to Shapiro‐Wilk normality test (UNIVARIATE procedure; SAS 9.0 software, SAS Institute Inc., USA). Statistical significances of variables determined in the tissues were assessed with Student‐Newman‐Keuls (SNK) multiple comparisons test ( p < 0.05) (Sousa et al, ).…”

“…For protein extraction and analysis, we used all ten parts of the ram epididymides ( Figure 1) of the 500μL protease inhibitor (Sigma Aldrich, EUA). Subsequently, the material was centrifuged (4°C, 10,000 g) and the supernatant, transferred to new tubes (Souza et al, 2014). Then acetone (1:9) was added, centrifuged (5 min., 1,000 g, 4°C) and the pellet was solubilized in 0.5 ml PBS (1x).…”