Responses of Sri Lankan indigenous goats and their Jamnapari crosses to artificial challenge with Haemonchus contortus.

Goat farming plays an important role in the Sri Lankan rural economy. Sri Lankan indigenous (SLI) goats and their crossbreds are reared mainly under extensive management and indiscriminately exposed to pathogens and parasites. This study was designed to evaluate resistance to haemonchosis in SLI goats and their Jamnapari crossbreds (JCB) in the dry zone of Sri Lanka. Twenty SLI and 20 JCB 4-month-old male goats were artificially challenged with 5000 H. contortus L3 larvae. Faecal egg counts (FEC), body weights, FAffa MAlan CHArt (FAMACHA®) scores, packed cell volumes (PCV), red blood cell counts, total and differential white blood cell counts, blood haemoglobin contents, serum total protein and albumin contents, and serum pepsinogen and antibody levels were determined at 0, 21, 28, 35 and 42days after challenge. Effects of measurement time were significant for all variables (P<0.05). Breed effects approached significance (P=0.06) and measurement time×breed interaction was significant (P<0.05) for FEC. Peak FEC occurred at day 35 in both goat types, and JCB goats had higher FEC than SLI goats at days 28 (P<0.001), 35 (P<0.10), and 42 (P<0.10). Means for FEC at day 35 were 1783±446 eggs per gram of feces (epg) for SLI kids and 3329±850 epg for JCB kids. Haematological parameters, serum chemistry, and FAMACHA scores suggested that SLI goats were recovering from parasitic infection by day 42, whereas JCB goats had increasing severity of anaemia. Means for PCV in SLI goats decreased from 26.8±0.7% at day 0 to 19.7±0.9% at day 35 and thereafter increased to 20.2±0.9% at day 42. Means for PCV in JCB goats declined from 25.9±0.6% at day 0 to 17.2±0.9% at day 42. Eosinophilia was observed in both genotypes. The JCB goats were heavier than SLI goats and had higher antibody titres, reflecting higher levels of parasitism. Both goat types significantly increased in body weight during the experiment and therefore tolerated parasite infection without severe production losses. We concluded that SLI goats were more resistant to haemonchosis than JCB goats, but that JCB goats were somewhat resilient to parasitic infection. Substantial variability in measurements associated with parasite infection in both breeds indicated potential to improve parasite resistance. Phenotypic information should be coupled with genomic information to identify appropriate breeding goals for future selection programs.

Thyroid hormone and anti-Mullerian hormone (AMH) on Leydig cell differentiation: studies using C57BL/6 mice and AMH over expressing mice.

Although the thyroid hormone has stimulatory effects and anti-Mullerian hormone (AMH) has inhibitory effects on prepubertal Leydig cell (LC) differentiation, it is important to find out whether the stimulatory effect of thyroid hormone could overcome the inhibitory effect of AMH on postnatal LC differentiation. Therefore, the objective of the present study was to use the anti-Mullerian hormone overexpressing mouse (AMH++) model to understand the simultaneous effects of AMH and thyroid hormone on postnatal LC differentiation, proliferation, maturation and function and to test whether the inhibitory effect of AMH could be overcome by the stimulatory effect of the thyroid hormone. Four age groups (7, 21, 40, 90 days) of control (C57BL/6; C) and AMH++ were used. Mice received either saline or triiodothyronine (T3) SC injections daily from birth to 21days. The four experimental groups were C, C+T3, AMH++ and AMH+T3. Body and testis weights of both C+T3 and AMH+T3 mice were significantly reduced at days 21, 40 and 90, compared to their age-matched saline-treated mice (C and AMH++). BrdU studies revealed the absence of LC proliferation in AMH++ mice at day7, however, same-aged mice of C+T3 and AMH+T3 mice showed increased LC proliferation; the rate was highest in C+T3 at day21. C+T3 mice of day 21 had more LC than C mice as well as AMH+T3 and AMH++ mice. At days 40 and 90, LC number/testis in C+T3 was lower than C, however, AMH+T3 had higher LC numbers than AMH++ mice. Cellular apoptosis was not seen as the cause of reduced LC numbers. Serum testosterone was not different among groups at day 21, but significantly higher levels were seen in AMH+T3 compared to AMH++ mice at days 40 and 90. Similar pattern was seen for luteinizing hormone (LH)-stimulated testicular testosterone and androstenedione production in vitro. Findings suggest that T3-treatment for the first postnatal 21 days was able to partially counteract the inhibitory effect of AMH on prepubertal LC differentiation. Whether continuation of the T3-treatment beyond 21 days would have resulted in complete removal of this inhibition, is a question that needs to be addressed.

Expression of insulin-like peptide 3 in the postnatal rat Leydig cell lineage: timing and effects of triiodothyronine-treatment.

Fetal (FLC) and adult Leydig cells (ALC) secrete insulin-like peptide 3 (INSL3), which is linked to cryptorchidism in the newborn rat. Its gene regulation appears to be independent of that for most steroidogenic enzymes, and may thus be a marker for other aspects of ALC differentiation. Our study examined the following on INSL3 peptide expression in ALC lineage (i) timing, (ii) which cell stage, and (iii) effects of triiodothyronine (T3). Male Sprague-Dawley (SD) rats of postnatal days (pd) 1, 5, 7-21, 28, 40, 60, and 90 were used for the objectives (i) and (ii). For the objective (iii), control and T3-treated (daily T3 SC, 50 mug/kg bw) SD rats of pd7-16 and 21 were used. INSL3 was immunolocalized in Bouin's-fixed testes. FLC were positive and mesenchymal and Leydig progenitor cells were negative for INSL3 at tested ages. INSL3 in ALC lineage was first detected in newly formed ALC on pd16, although they were present from pd10. The intensity of INSL3 label was greater in ALC of pd40-90. ALC were present in T3-treated testes at pd9, but INSL3 first detected in them was on pd12. While INSL3 in FLC regulates testicular descent, INSL3 in ALC still has no well-defined function. However, its pattern of expression correlates temporally with the development of steroidogenic function and spermatogenesis. Thus, the delay between ALC differentiation and INSL3 expression in them implies that INSL3 in ALC is associated with maturation. The advancement of INSL3 expression in the ALC of T3-treated rats implies that this function is established earlier with T3-treatment.

Detection of anti-Mullerian hormone receptor II protein in the postnatal rat testis from birth to sexual maturity.

Anti-Mullerian hormone (AMH) produced by the immature Sertoli cells negatively regulates the postnatal Leydig cell (i.e. adult Leydig cells/ALC) differentiation, however, the mechanism is sparsely understood. AMH negatively regulates the steroidogenic function of fetal Leydig cells (FLC) and ALC. However, when this function is established in the ALC lineage and whether AMH has a function in FLC in the postnatal testis are not known. Therefore, the objectives of this study were to examine the presence of AMH receptor type II (AMHR-II) in FLC and cells in the ALC lineage in the postnatal mammalian testis using the rat model Male Sprague Dawley rats of days 1, 5, 7-21, 28, 40, 60 and 90 were used. AMHR-II in testicular interstitial cells was detected in testis tissue using immunocytochemistry. Findings showed that the mesenchymal and the progenitor cells of the ALC lineage, were negative for AMHR-II. The newly formed ALC were the first cell type of the ALC lineage to show positive labeling for AMHR-II, and the first detection was on postnatal day 13, although they were present in the testis from day 10. From days 13-28, labeling intensity for AMHR-II in the ALC was much weaker than those at days 40-90. FLC were also positive. The time lag between the first detection of the newly formed ALC in the testis and the first detection of AMHR-II in them suggests that the establishment of the negative regulatory role of AMH on ALC steroidogenesis does not take place immediately upon their differentiation; no change in cell size occurs during this period. The absence of AMHR-II in mesenchymal cells suggests that it is unlikely that the negative regulatory effect of AMH on ALC differentiation in the postnatal testis is achieved via a direct action of AMH on mesenchymal cells. The presence of AMHR-II in postnatal FLC suggests a possible role by AMH on FLC, which warrants future investigations.

Effects of thyroid hormones on Leydig cells in the postnatal testis.

Thyroid hormones (TH) stimulate oxidative metabolism in many tissues in the body, but testis is not one of them. Therefore, in this sense, testis is not considered as a target organ for TH. However, recent findings clearly show that TH have significant functions on the testis in general, and Leydig cells in particular; this begins from the onset of their differentiation through aging. Some of these functions include triggering the Leydig stem cells to differentiate, producing increased numbers of Leydig cells during differentiation by causing proliferation of Leydig stem cells and progenitors, stimulation of the Leydig cell steroidogenic function and cellular maintenance. The mechanism of action of TH on Leydig cell differentiation is still not clear and needs to be determined in future studies. However, some information on the mechanisms of TH action on Leydig cell steroidogenesis is available. TH acutely stimulate testosterone production by the Leydig cells in vitro via stimulating the production of steroidogenic acute regulatory protein (StAR) and StAR mRNA in Leydig cells; StAR is associated with intracellular trafficking of cholesterol into the mitochondria during steroid hormone synthesis. However, the presence and/or the types of TH receptors in Leydig cells and other cell types of the Leydig cell lineage is still to be resolved. Additionally, it has been shown that thyrotropin-releasing hormone (TRH), TRH receptor and TRH mRNA in the testis in many mammalian species are seen exclusively in Leydig cells. Although the significance of the latter observations are yet to be determined, these findings prompt whether hypothalamo-pituitary-thyroid axis and hypothalamo-pituitary-testis axis are short-looped through Leydig cells.