Silva M, Fernández A, Ulloa-Leal C, Adams G P, Berland M A, Ratto M H
Escuela de Medicina Veterinaria, Facultad de Recursos Naturales, Universidad Católica de Temuco, Temuco, Chile.
Department of Animal Science, Faculty of Veterinary Sciences, Universidad Austral de Chile, Valdivia, Chile.
Theriogenology. 2015 Oct 15;84(7):1096-102. doi: 10.1016/j.theriogenology.2015.06.006. Epub 2015 Jun 20.
The objective of the study was to compare the pituitary and ovarian responses after intramuscular, intravenous, or intrauterine administration of β-nerve growth factor (β-NGF) of seminal plasma origin (SP-NGF) in llamas. In experiment 1, mature female llamas with a growing follicle of 7 mm or greater were assigned randomly to four groups (n = 7/group) and given 2 mg of purified SP-NGF in a volume of 2 mL by (1) intramuscular administration, (2) intravenous administration, and (3) intrauterine infusion, or (4) intrauterine infusion of 2 mL of PBS (negative control). Because ovulations were not detected after intrauterine infusion in experiment 1, a second experiment was done to determine if a higher dose of SP-NGF given by intrauterine infusion, similar to a natural dose during copulation, will elicit an ovulatory response. In experiment 2, llamas with a growing follicle of 7 mm or greater were assigned randomly to three groups (n = 6/per group) given an intrauterine infusion of (1) 4 mL of raw seminal plasma, (2) 4 mL of PBS containing 20 mg of purified llama SP-NGF, or 3) 4 mL of PBS (negative control). In both experiments, the ovaries were examined daily by transrectal ultrasonography using a B-mode scanner and power Doppler mode to detect ovulation and to monitor CL growth, regression, and vascularization. Blood samples were collected to determine plasma LH and progesterone concentrations. In experiment 1, only llamas treated by intramuscular or intravenous administration of SP-NGF ovulated (7 of 7 and 6 of 7, respectively). Plasma LH concentration did not differ between the intramuscular and intravenous SP-NGF-treated groups, nor did CL diameter, CL vascularization, or plasma progesterone concentration profiles. In experiment 2, the ovulation rate was 100% for llamas treated by intrauterine infusion of raw seminal plasma or llama SP-NFG, whereas no ovulations were detected in females treated with PBS. Plasma LH concentrations did not differ between groups that ovulated, nor did CL diameter, CL vascularization, or plasma progesterone concentration profiles. We conclude that β-NGF from llama seminal plasma origin elicits a preovulatory LH surge, followed by ovulation and the development of a functional CL, regardless of the route of administration. However, the dose required to elicit pituitary and ovarian responses is higher when administered by intrauterine infusion than by intramuscular or intravenous routes.
本研究的目的是比较肌肉注射、静脉注射或子宫内注射源于精浆的β-神经生长因子(β-NGF)后,美洲驼垂体和卵巢的反应。在实验1中,将有一个7毫米或更大生长卵泡的成年雌性美洲驼随机分为四组(每组n = 7),并通过以下方式给予2毫克纯化的精浆β-NGF(SP-NGF),体积为2毫升:(1)肌肉注射,(2)静脉注射,(3)子宫内输注,或(4)子宫内输注2毫升磷酸盐缓冲液(PBS,阴性对照)。由于在实验1的子宫内输注后未检测到排卵,因此进行了第二个实验,以确定子宫内输注更高剂量的SP-NGF(类似于交配期间的自然剂量)是否会引发排卵反应。在实验2中,将有一个7毫米或更大生长卵泡的美洲驼随机分为三组(每组n = 6),进行子宫内输注:(1)4毫升未加工的精浆,(2)4毫升含有20毫克纯化美洲驼SP-NGF的PBS,或(3)4毫升PBS(阴性对照)。在两个实验中,每天使用B型扫描仪和功率多普勒模式经直肠超声检查卵巢,以检测排卵并监测黄体(CL)的生长、退化和血管形成。采集血样以测定血浆促黄体生成素(LH)和孕酮浓度。在实验1中,仅通过肌肉注射或静脉注射SP-NGF处理的美洲驼排卵(分别为7只中的7只和7只中的6只)。肌肉注射和静脉注射SP-NGF处理组之间的血浆LH浓度没有差异,CL直径、CL血管形成或血浆孕酮浓度曲线也没有差异。在实验2中,子宫内输注未加工精浆或美洲驼SP-NFG处理的美洲驼排卵率为100%,而用PBS处理的雌性未检测到排卵。排卵组之间的血浆LH浓度没有差异,CL直径、CL血管形成或血浆孕酮浓度曲线也没有差异。我们得出结论,源于美洲驼精浆的β-NGF会引发排卵前LH激增,随后排卵并形成功能性黄体,无论给药途径如何。然而,通过子宫内输注给药时,引发垂体和卵巢反应所需的剂量高于肌肉注射或静脉注射途径。