Hormone-sensitive lipase deficiency affects the expression of SR-BI, LDLr, and ABCA1 receptors/transporters involved in cellular cholesterol uptake and efflux and disturbs fertility in mouse testis.

Hormone-sensitive lipase (HSL) hydrolyse acylglycerols, cholesteryl and retinyl esters. HSL is a key lipase in mice testis, as HSL deficiency results in male sterility. The present work study the effects of the deficiency and lack of HSL on the localization and expression of SR-BI, LDLr, and ABCA1 receptors/transporters involved in uptake and efflux of cholesterol in mice testis, to determine the impact of HSL gene dosage on testis morphology, lipid homeostasis and fertility. The results of this work show that the lack of HSL in mice alters testis morphology and spermatogenesis, decreasing sperm counts, sperm motility and increasing the amount of Leydig cells and lipid droplets. They also show that there are differences in the localization of HSL, SR-BI, LDLr and ABCA1 in HSL+/+, HSL+/- and HSL-/- mice. The deficiency or lack of HSL has effects on protein and mRNA expression of genes involved in lipid metabolisms in mouse testis. HSL-/- testis have augmented expression of SR-BI, LDLr, ABCA1 and LXRß, a critical sterol sensor that regulate multiple genes involved in lipid metabolism; whereas LDLr expression decreased in HSL+/- mice. Plin2, Abca1 and Ldlr mRNA levels increased; and LXRα (Nr1h3) and LXRß (Nr1h2) decreased in testis from HSL-/- compared with HSL+/+; with no differences in Scarb1. Together these data suggest that HSL deficiency or lack in mice testis induces lipid homeostasis alterations that affect the cellular localization and expression of key receptors/transporter involved in cellular cholesterol uptake and efflux (SR-BI, LDRr, ABCA1); alters normal cellular function and impact fertility.

Lack of Improvement of Sperm Characteristics in Obese Males After Obesity Surgery Despite the Beneficial Changes Observed in Reproductive Hormones.

BACKGROUND: Even though obesity surgery normalizes circulating testosterone concentrations in males with obesity-associated secondary hypogonadism, its impact on spermatogenesis remains controversial. We aimed to evaluate sperm characteristics in obese men after bariatric surgery as well as changes in reproductive hormones. METHODS: Twenty severely obese men (body mass index (BMI) ≥ 35 kg/m2) were evaluated before and 2 years after bariatric surgery. The serum was assayed for insulin, leptin, kisspeptin, and inhibin B, among other hormones. Homeostasis model assessment of insulin resistance (HOMA-IR) was estimated. We used World Health Organization reference values for sperm analysis. RESULTS: After surgery, serum total testosterone, calculated free testosterone, inhibin B, and kisspeptin increased, whereas fasting insulin, HOMA-IR, and leptin concentrations decreased. Despite these improvements, sperm volume showed a small decrease after surgery, while the rest of sperm characteristics remained mostly unchanged. Abnormal sperm concentration persisted in 60% of the patients. CONCLUSIONS: Sperm characteristics may not improve after bariatric surgery despite the beneficial changes of reproductive hormones.

Morbid obesity-related changes in the expression of lipid receptors, transporters, and HSL in human sperm.

OBJECTIVE: To study the location and expression of receptors (SR-BI/CLA-1, SR-BII, and LDLr) and transporter (ABCA1) involved in uptake and efflux of cholesterol in human spermatozoa and assess whether obesity alters its location/expression and whether this could be related to infertility. DESIGN: Observational study. SETTING: None PATIENT(S): Ten controls and 20 obese patients. INTERVENTION(S): Anthropometric parameters. Serum and semen samples were collected. MAIN OUTCOME MEASURE(S): Spermatozoon concentration, immunolocalization, and protein expression in semen. RESULTS: Spermatozoon concentration and motility was decreased in morbidly obese patients. SR-BI/CLA-1, SR-BII, LDLr, and ABCA1 are located in the spermatozoon cell membrane and the localization does not change between obese patients and controls. Control spermatozoa showed high SR-BI expression, and less expression for the rest of the receptors analyzed, indicating that SR-BI/CLA-1 is relevant in human spermatozoon cholesterol uptake/efflux. On the contrary, spermatozoa of obese patients showed less SR-BI/CLA-1 expression than controls, and more intense positive staining for SR-BII, LDLr, and ABCA1. Finally, human sperm expresses the 130- and 82-kDa hormone-sensitive lipase (HSL) isoforms. The 130-kDa isoform is expressed in the control sperm, and the expression disappears in the obese patients. CONCLUSION(S): The presence of lipid receptors/transporters and HSL in human spermatozoa suggests their role in the process of maturation/capacitation. The changes in the expression of lipid receptors/transporters and the lack of the 130-kDa HSL isoform in obese patients prevent the hydrolysis of cholesterol esters internalized by these receptors, and favor their accumulation in the cytoplasm of the spermatozoa that could contribute to lipotoxicity and infertility.

HSL-knockout mouse testis exhibits class B scavenger receptor upregulation and disrupted lipid raft microdomains.

There is a tight relationship between fertility and changes in cholesterol metabolism during spermatogenesis. In the testis, class B scavenger receptors (SR-B) SR-BI, SR-BII, and LIMP II mediate the selective uptake of cholesterol esters from HDL, which are hydrolyzed to unesterified cholesterol by hormone-sensitive lipase (HSL). HSL is critical because HSL knockout (KO) male mice are sterile. The aim of the present work was to determine the effects of the lack of HSL in testis on the expression of SR-B, lipid raft composition, and related cell signaling pathways. HSL-KO mouse testis presented altered spermatogenesis associated with decreased sperm counts, sperm motility, and infertility. In wild-type (WT) testis, HSL is expressed in elongated spermatids; SR-BI, in Leydig cells and spermatids; SR-BII, in spermatocytes and spermatids but not in Leydig cells; and LIMP II, in Sertoli and Leydig cells. HSL knockout male mice have increased expression of class B scavenger receptors, disrupted caveolin-1 localization in lipid raft plasma membrane microdomains, and activated phospho-ERK, phospho-AKT, and phospho-SRC in the testis, suggesting that class B scavenger receptors are involved in cholesterol ester uptake for steroidogenesis and spermatogenesis in the testis.

Efecto de la dosis de eCG sobre la inducción del celo en cabras mestizas luego de un tratamiento corto con Medroxiprogesterona / Effect of eCG Dosage on Estrus Induction in Crossbred Goats After a Short-Term Medroxiprogesterone Treatment

El propósito del presente estudio fue evaluar el efecto de la dosis de gonadotropina coriónica equina (eCG) sobre la inducción del celo en cabras mestizas manejadas bajo condiciones tropicales. Las cabras (n = 32) fueron tratadas con una esponja intravaginal impregnada con 50 mg de acetato de medroxiprogesterona (MAP) durante seis días. Al momento del retiro de las esponjas, las cabras fueron divididas al azar en tres grupos: control (GC, n=11), no recibieron eCG; G250: (n=10) recibieron 250 UI de eCG vía intramuscular y G500: (n=11) recibieron 500 UI de eCG vía intramuscular. La eCG no afectó la tasa de expresión de celo, siendo 81,8% (9/11) para el GC y de 100% para los grupos G250 y G500 (P>0,05). El intervalo al celo fue significativamente más corto en el grupo G500 (32,54 ± 3,00 horas, P<0,05) que en los grupos G250 y GC (40,65 ± 3,27 y 42,20 ± 3,54 horas, respectivamente). La duración del celo no difirió entre los grupos (P>0,05). El porcentaje de cabras repitiendo celo, luego de un ciclo estrual de corta duración, varió según el grupo, siendo de 27,3% (3/11) en el grupo G500 y de 10% (1/10) en el grupo G250; mientras que ninguna cabra del grupo control repitió celo luego de un ciclo estrual corto; no se observaron diferencias significativas entre los grupos (P>0,05). En conclusión, la eCG no es necesaria para inducir el celo en cabras tratadas por seis días con una esponja intravaginal conteniendo MAP, sin embargo, la dosis de 500 UI de eCG fue necesaria para acortar el intervalo al celo.

The aim of this study was to evaluate the effect of dosage of equine chorionic gonadotrophin (eCG) dose on the estrus induction after a short-term medroxyprogesterone treatment in crossbred goat under tropical conditions. Goats (n = 32) received an intravaginal sponge containing 50 mg of medroxyprogesterone acetate (MAP) for six days, at time of sponge withdrawal goats were divided randomly in three groups: CG, control group (n=11), did not received eCG; the second group (G250, n=10), received 250 IU of eCG intramuscular and the third group (G500, n =11), received 500 IU of eCG IM. Treatment with eCG did not affect the rate of estrus presentation, being 81.8% (9/11) for GC group and 100% for groups G250 (10/10) and G500 (11/11), P>0.05. The interval to estrus was significantly shorter in group G500 (32.54 ± 3.00 hours, P<0.05) than GC and G250 (40.65 ± 3.27 y 42.20 ± 3.54 hours, respectively). Estrus duration was not affected by eCG treatment (P>0.05). A shorter interestrus interval was observed in 27.3% (3/11) of goats in G500 and 10% (1/10) of goats in G250, while no goats in GC had a shorter interestrus interval. In conclusion, eCG is not necessary to induce estrus in goats treated for six days with an intragavinal sponges containing MAP, however the dose of 500 IU was necessary to shortens the interval to the estrus.

Hormone-sensitive lipase expression and IHC localization in the rat ovary, oviduct, and uterus.

Hormone-sensitive lipase (HSL) is a key regulator of cholesterol esters metabolism. The aim of this study was to determine HSL localization in rat female reproductive organs during the ovarian cycle by IHC methods. HSL was located in the ovarian epithelium. The granulosa cells and oocytes of primordial follicles were immunonegative. In mature follicles, HSL was found in oocytes and theca and granulosa cells. However, HSL expression in theca cells and oocytes decreased during follicular atresia. Luteal cells showed HSL staining in cytoplasm during proestrus and estrus, in the nucleus during metestrus, and in cytoplasm and the nucleus during diestrus. In the tubaric ampulla, HSL was located in the epithelial cells nuclei and in the cilia during proestrus and estrus but mainly in the nucleus during metestrus and diestrus. In the isthmus, cells showed HSL immunolabeling in the nucleus and cilia during proestrus, but only in the cilia during estrus, metestrus, and diestrus. In the uterus, HSL was found in the epithelial cells nuclei. HSL-immunoreactive bands at 84, 67, 54, and 43 kDa were found in rat female reproductive organs. HSL labeling in the nucleus of epithelial and germ cells suggests an as yet unknown function for this protein, probably related to oogenesis and cell proliferation.

Brief communication: effect of post-mating progestagen administration on pregnancy rate in crossbred goats following an induced estrus

Los celos inducidos con progesterona o progestagenos son de menor fertilidad en comparación con los celos naturales. Sin embargo, la administración postmonta de progesterona ha sido reportada por incrementar el desarrollo embrionario y la tasa de preñez; por lo tanto, el propósito de este estudio fue evaluar el efecto de la administración postmonta de un progestageno sobre la tasa de preñez en cabras mestizas tropicales luego de un celo inducido con acetato de medroxiprogestrona (MAP) durante el periodo temprano postparto. La inducción del celo se inicio el día 31,3 ± 1,7 postparto mediante la utilización de esponjas intravaginales impregnadas con 60 mg de MAP durante 14 días y al momento del retiro de las esponjas se aplicaron 500 UI de eCG, vía intramuscular. El celo fue detectado cada seis horas a partir de 24 horas de retiradas las esponjas. El servicio se realizó por monta natural con dos machos enteros. Cinco días luego de la monta, las cabras servidas fueron distribuidas de forma aleatoria en dos grupos, el grupo control (n=18), sin tratamiento; y el grupo MAP (n=18), que recibió una esponja con 60 mg de MAP por 14 días. El diagnóstico ultrasonográfico de preñez se realizó a los 50 días postmonta. El tratamiento con MAP no afectó la tasa de preñez, con 44,4 por ciento (8/18) en el grupo MAP, mientras que en el grupo control fue de 55,5 por ciento (10/18), P>0,05. En conclusión, el tratamiento con una esponja impregnadas con 60 mg de MAP entre los días 5 y 19 postmonta no afectó la tasa de preñez luego de un celo inducido durante el período postparto temprano en cabras mestizas tropicales.

Estrus induced with progesterone or progestagens have low fertility compared to natural estrus. However, post-mating progesterone administration has been reported by increases embryo development and pregnancy rate; therefore the aim of this study was to evaluate the effect of post-mating progestagen administration on pregnancy rate after medroxyprogesterone acetate (MAP) induced estrus in crossbred goats. Estrus induction was started at 31.3 ± 1.7 days post-partum with intra- vaginal sponges impregnated with 60 mg of MAP during 14 days and at time of sponge removal were applied 500 IU of eCG intramuscular. Estrus was detected every 6 hours from 24 hours onwards after sponge withdrawal. Goats were naturally mated with two entry bucks. Five days post-mating, mated goats were randomly assorted into two groups, control group (n=18), without any treatment, and MAP group (n=18), receiving a sponge with 60 mg of MAP for fourteen days. Ultrasonographyc diagnosis of pregnancy was performed at day 50 post-mating. MAP treatment, did not affect the pregnancy rate, with 44.4% (8/18) in MAP group, while in control group was 55.5% (10/18), P>0.05. In conclusion, MAP treatment with a sponge impregnated with 60 mg of MAP between days 5 and 19 post-mating did not affect the pregnancy rate after progestagen induced estrus during the early post-partum period in crossbred tropical goats.

Expression, activity, and localization of hormone-sensitive lipase in rat mammary gland during pregnancy and lactation.

We examined the presence of hormone-sensitive lipase (HSL) in mammary glands of virgin, pregnant (12, 20, and 21 days), and lactating (1 and 4 days postpartum) rats. Immunohistochemistry with antibody against rat HSL revealed positive HSL in the cytoplasm of both alveolar epithelial cells and adipocytes. In virgin rats, immunoreactive HSL was observed in mammary adipocytes, whereas diffuse staining was found in the epithelial cells. Positive staining for HSL was seen in the two types of cells in pregnant and lactating rats. However, as pregnancy advanced, the staining intensity of immunoreactive HSL increased in the epithelial cells parallel to their proliferation, attaining the maximum during lactation. An immunoreactive protein of 84 kDa and a HSL mRNA of 3.3. kb were found in the rat mammary gland as in white adipose tissue. Both HSL protein and activity were lower in mammary glands from 20 and 21 day pregnant rats than from those of virgin rats, although they returned to virgin values on days 1 and 4 of lactation. Mammary gland HSL activity correlated negatively to plasma insulin levels. Immunoreactive HSL and HSL activity were found in lactating rats' milk. The observed changes indicate an active role of HSL in mammary gland lipid metabolism.