Bovine reaginic antibody III. Cross-reaction of antihuman IgE and antibovine reaginic immunoglobulin antisera with sera from several species of mammals.

Using antisera specific for the heavy chain of human IgE and bovine reaginic immunoglobulin, the degree of cross-reaction amongst sera from pig, rat, rabbit, guinea pig, goat, cow, horse, dog, cat and human was tested. Antihuman IgE antiserum gave strong reactions with pig, rabbit, cow, goat and human sera (100% to 15.1%) and weak reactions with rat, guinea pig, horse, dog and cat sera (10.1% to 3.22%). Antibovine reagin antiserum produced a considerable amount of cross-reaction with sera from pig, rat, rabbit, goat, horse and human (43.6% to 20.1%) with limited reactions with guinea pig, dog and cat sera (13.9% to 9.3%).     

Angiostatin and integrin alphavbeta3 in the feline, bovine, canine, equine, porcine and murine retina and cornea

PURPOSE: Angiogenesis is tightly controlled in the ocular tissues of domestic animals but its mechanisms are not fully understood. This is largely because of insufficient data on the expression of molecules that impact angiogenesis. Because angiostatin and one of its receptors integrin alphavbeta3 inhibit and promote angiogenesis, respectively, we hypothesized that the normal retina and cornea of domestic animals would express angiostatin but not integrin alphavbeta3. PROCEDURE: Normal eyes of the cat, cow, dog, horse, pig and rat were evaluated for angiostatin and integrin alphavbeta3 by light and electron immunocytochemistry and estern blots. RESULTS: Angiostatin was detected in the corneal epithelium of the cat, dog, horse, pig and rat, but was not found in cow corneal epithelium. Angiostatin was localized in the nerve fiber layer, ganglion cell layer, inner and outer plexiform layers, and the photoreceptor layer of the cat, cow, dog and rat. Horse and pig retinas showed additional staining in the matrix of the inner nuclear layer. Immunogold electron microscopy further confirmed angiostatin in cat retina. Western blots showed angiostatin in corneal and retinal homogenates. Integrin alphavbeta3 was absent in cornea and retina of all the species studied. CONCLUSION: These data show that angiostatin, an inhibitor of angiogenesis, is present while integrin alphavbeta3, which promotes angiogenesis, is absent in normal cornea and retina of the domestic animals in this study with the exception being angiostatin absence in cow corneal epithelium. Therefore, angiostatin may contribute to the anti-angiogenic environment in the normal domestic animal eye while its absence in the cow may contribute to greater propensity for corneal vascularization. Because integrin alphavbeta3 is one of the receptors for angiostatin, its absence may prevent angiostatin from killing normal retinal and corneal cells.     

Responses of seasonally anovulatory mares to daily administration of thyrotropin-releasing hormone and(or) gonadotropin-releasing hormone analog

Seventeen seasonally anovulatory light horse mares were treated daily, starting January 5 (d 1), for 28 d with GnRH analog (GnRH-A; 50 ng/kg BW) and(or) thyrotropin-releasing hormone (TRH; 5 microg/kg BW) in a 2 x 2 factorial arrangement of treatments to test the hypothesis that combined treatment may stimulate follicular growth and development. Ovaries were examined via ultrasonography and jugular blood samples were collected every 3 d. Frequent blood samples were collected after treatment injections on d 1, 2, 4, 7, 11, 16, and 22; on d 29, all mares received an i.v. mixture of GnRH, TRH, sulpiride, and EP51389 (a growth hormone secretagogue) to assess pituitary responsiveness. No consistent effects (P > 0.1) of treatment were observed for plasma LH, FSH, prolactin, or thyroxine concentrations in samples collected every 3 d. The only effect on ovarian follicle numbers was a reduction in number of follicles 11 to 19 mm in diameter due to TRH treatment (P = 0.029). No mare ovulated during treatment. On the days of frequent sampling, mean LH (P = 0.0001) and FSH (P = 0.001) concentrations were higher in mares receiving GnRH-A and tended to increase from d 1 through 7. In contrast, mean prolactin (P = 0.001) and thyroid-stimulating hormone (P = 0.0001) concentrations were high in mares receiving TRH on d 1 but rapidly decreased thereafter. When mares were administered the secretagogue mixture on d 29, the LH response was greater (P = 0.0002) in mares that had previously received GnRH-A but the FSH response was not affected (P > 0.1); the prolactin response was greater (P = 0.014) and the TSH response was smaller (P = 0.0005) in mares that had previously received TRH. Surprisingly, an immediate growth hormone response to EP51389 was absent in all mares. In conclusion, daily GnRH-A treatment stimulated plasma LH and FSH concentrations immediately after injection; although no long-term elevation in preinjection concentrations was achieved, the responses gradually increased over time, indicating a stimulation of gonadotropin production and storage. Daily treatment with TRH stimulated plasma TSH and prolactin concentrations, but the response diminished rapidly and was minimal within a few days, indicating a depletion of pituitary stores and little or no stimulation of production. There was no beneficial effect of adding TRH treatment to the daily GnRH-A regimen.     

Pituitary receptors for GnRH and estradiol, and pituitary content of gonadotropins in beef cows. II. Changes during the postpartum period

Pregnant beef heifers (n = 24) were assigned randomly to four groups and slaughtered at day 1, 15, 30 or 45 postpartum. The day prior to slaughter blood samples were taken from each cow every 15 min for 8 hr. The anterior pituitary gland, preoptic area (POA) and medial basal hypothalamus (HYP) were collected from each cow. Contents of gonadotropin-releasing hormone (GnRH) in extracts of POA and HYP, and luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in extracts of anterior pituitary were quantified by radioimmunoassay. In the anterior pituitary gland, membrane receptors for GnRH were quantified by a standard curve technique and cytosolic receptors for estradiol were quantified by saturation analysis. Concentrations of LH, FSH and prolactin in serum were quantified by radioimmunoassay. Only one cow of eight had a pulse of LH during the 8 hr bleeding period on day 1 postpartum. This increased to 8 pulses in 6 cows on day 30 postpartum. Contents of GnRH in POA (15.0 +/- 3.2 ng) and HYP (14.0 +/- 2.0 ng) did not change significantly during the postpartum period. Pituitary content of LH was low following parturition (.2 +/- .1 mg/pituitary) and increased significantly through day 30 postpartum (1.2 +/- .1 mg/pituitary). Pituitary content of FSH did not change over the postpartum period. Receptors for both GnRH (.9 +/- .2 pmoles/pituitary) and estradiol (5.0 +/- .9/moles/pituitary) were elevated on day 15 postpartum, possibly increasing the sensitivity of the anterior pituitary gland to these hormones and leading to an increased rate of synthesis of LH that restored pituitary content to normal by day 30 postpartum.     

Combined pituitary hormone deficiency in german shepherd dogs with dwarfism

In German shepherd dogs pituitary dwarfism is known as an autosomal recessive inherited abnormality. To investigate whether the function of cells other than the somatotropes may also be impaired in this disease, the secretory capacity of the pituitary anterior lobe (AL) cells was studied by a combined pituitary AL stimulation test with four releasing hormones (4RH test) in four male and four female German shepherd dwarfs. In addition, the morphology of the pituitary was investigated by computed tomography. The physical features of the eight German shepherd dwarfs were primarily characterized by growth retardation and stagnant development of the hair coat. The results of the 4RH test confirmed the presence of hyposomatotropism. The basal plasma TSH and prolactin concentrations were also low and did not change upon stimulation. Basal plasma concentrations of LH were relatively low and responded only slightly to suprapituitary stimulation. With respect to the plasma FSH levels there was a clear gender difference. In the males plasma FSH concentrations remained below the detection limit throughout the 4RH test, whereas in the females the basal plasma FSH levels were slightly lower and there was only a small increase following suprapituitary stimulation, compared with the values in age-matched controls. In contrast, basal and stimulated plasma ACTH concentrations did not differ between the dwarfs and the controls. Computed tomography of the pituitary fossa revealed a normal sized pituitary with cysts in five dogs, an enlarged pituitary with cysts in two dogs, and a small pituitary gland without cysts in the remaining dog. The results of this study demonstrate that German shepherd dwarfs have a combined deficiency of GH, TSH, and prolactin together with impaired release of gonadotropins, whereas ACTH secretion is preserved. The combined pituitary hormone deficiency is associated with cyst formation and pituitary hypoplasia.     

Effect of LPS on Reproductive System at the Level of the Pituitary of Anestrous Ewes

In our research we focused our attention on the effect of the immune stress induced by bacterial endotoxin–lipopolysaccharide (LPS) on the hypothalamic–pituitary–gonadal axis (HPG) at the pituitary level. We examined the effect of intravenous (i.v.) LPS injection on luteinizing hormone (LH) and follicle‐stimulating hormone (FSH) release from the anterior pituitary gland (AP) in anestrous ewes. The effect of endotoxin on prolactin and cortisol circulating levels was also determined. We also researched the effect of immune challenge on the previously mentioned pituitary hormones and their receptors genes expression in the AP. Our results demonstrate that i.v. LPS injection decreased the plasma concentration of LH (23%; p < 0.05) and stimulates cortisol (245%; p < 0.05) and prolactin (60%; p < 0.05) release but has no significant effect on the FSH release assayed during 6 h after LPS treatment in comparison with the control levels. The LPS administration affected the genes expression of gonadotropins’β‐subunits, prolactin and their receptors in the AP. Endotoxin injection significantly decreased the LHβ and LH receptor (LHR) gene expression (60%, 64%; p < 0.01 respectively), increased the amount of mRNA encoding FSHβ, FSH receptor (FSHR) (124%, 0.05; 166%, p < 0.01; respectively), prolactin and prolactin receptor (PRLR) (50%, 47%, p < 0.01; respectively). The presented, results suggest that immune stress is a powerful modulator of the HPG axis at the pituitary level. The changes in LH secretion could be an effect of the processes occurring in the hypothalamus. However, the direct effect of immune mediators, prolactin, cortisol and other components of the hypothalamic pituitary–adrenal (HPA) axis on the activity of gonadotropes has to be considered as well. Those molecules could affect LH synthesis directly through a modulation at all stages of LHβ secretion as well as indirectly influencing the GnRHR expression and leading to reduced pituitary responsiveness to GnRH stimulation.     

上海地区多种动物莱姆病的血清学调查

为探讨猪、牛、羊、马、犬、鸡、鸭和野鸟等与人类关系密切的多种动物莱姆病的血清流行病学状况,从上海地区采集血清870份,应用酶联荧光测定技术(ELFA)检测莱姆病抗体。猪、牛、羊、鸡、鸭和野鸟中均未检测到莱姆病抗体,马血清中莱姆病抗体阳性率为18.5%,犬血清中莱姆病抗体阳性率为12.3%。结果表明,上海地区动物群中莱姆病的感染率相对较低,马和犬在莱姆病传播中的重要性应引起重视。     

Responses of Adenohypophyseal Hormones to Substance P Administration in Geldings: Comparison to Responses After Brief Exercise and Sulpiride Administration

Nine adult geldings were used in three experiments to study the possible role of substance P in the prolactin responses to nondopaminergic stimuli. Experiment 1 was performed as an incomplete Latin square design to determine the secretory responses of prolactin, growth hormone (GH), adrenocorticotropic hormone (ACTH), thyroid-stimulating hormone (TSH), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) to IV administration of substance P. Doses tested and compared to no peptide (0 dose, control) were 62, 125, 250, and 500 μg of substance P. The three highest doses of peptide caused an immediate rise in heart rate, sweating, salivation, rhinorrhea, stretching of hind legs, and defecation. The lowest dose (62 μg) caused minor sweating, some rhinorrhea, and a rise in heart rate. Recovery from these physical responses was complete in approximately 30 minutes. All doses of substance P caused an immediate rise (P < .01) in plasma prolactin concentrations, with the three highest doses producing similar responses, and the 62 μg dose producing a minimal response (P < .05). Concentrations of ACTH (P < .01) and GH (P = .05) also increased after substance P administration; concentrations of LH, FSH, and TSH were unaffected. Experiment 2 compared the effects of brief exercise on hormonal characteristics. Two minutes of trotting increased (P < .01) plasma concentrations of GH, ACTH, and prolactin, as well as LH (P = .055). Experiment 3 determined the relative responses of prolactin to a fixed dose of sulpiride (0.1 mg/kg of body weight). In general, the prolactin responses to substance P were similar to those after exercise, which were both generally less than after sulpiride. These data are consistent with a possible role of substance P in the prolactin response to stressful stimuli.     

Concentrations of luteinizing hormone, follicle stimulating hormone and prolactin following transection of the pituitary stalk in ovariectomized ewes

Two experiments (Spring and Fall) were conducted in ovariectomized ewes to determine changes in pituitary hormone secretion immediately after pituitary stalk-transection. Ewes underwent either pituitary stalk-transection (SS), sham-transection (SH) or administration of anesthesia only (AO). Stalk-transected, but not sham-operated or anesthetized ewes had polyuria and polydipsia for 7 to 14 days after surgery. Concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH) and prolactin were measured in peripheral blood samples collected every 10 minutes for a six-hour period. Results were comparable for each season. During the six hours following surgery or removal from anesthesia, concentrations of LH declined in all ewes, but more slowly in SS ewes. No differences in patterns or mean concentrations of FSH were observed. Immediately after surgery, concentrations of prolactin were elevated, then declined in SH and SS ewes. The decrease was greater in SH than SS ewes. Data are consonant with the view that hypothalamic inhibition as well as LHRH stimulation regulate gonadotropin release by the pituitary.     

Immunocytochemical identification of Pit-1 containing cells in the anterior pituitary of hens

Our goal was to identify the cells expressing Pit-1 protein in chicken anterior pituitary. The anterior pituitaries were collected from laying hens after perfusion with formalin-PBS, and fixed with Bouin's fixative followed by paraffin embedding. Sections of the anterior pituitaries were immunostained for Pit-1 in the first staining sequence followed by staining for 6 types of pituitary hormones in the second sequence. Pit-1 positive nuclei were observed in the glandular cells in both the cephalic and caudal lobes. Pit-1 immunoreaction products were colocalized in the glandular cells immunopositive for growth hormone, thyroid-stimulating hormone, follicle-stimulating hormone, luteinizing hormone, adrenocorticotropic hormone or prolactin. These results indicate that Pit-1 protein induction occurs in 6 types of glandular cells, suggesting that Pit-1 may regulate hormone synthesis in each glandular cell in the chicken pituitary.     

长毛兔和獭兔白细胞介素-4基因的序列分析与真核表达

应用反转录—聚合酶链式反应（RT-PCR）技术,在国内首次从ConA诱导培养的长毛兔和獭兔外周血淋巴细胞总RNA中扩增出IL-4基因。将扩增基因克隆入pMD18-T载体,经序列测定分析表明,长毛兔和獭兔IL-4基因全长均为441 bp,两序列间碱基组成无差异（登录号：EF606761和EU639687）。长毛兔和獭兔IL-4基因编码147个氨基酸,其中前24个氨基酸组成信号肽,余下的123个氨基酸组成成熟肽。与已报道的犬、猫、人、猪、牛、羊、马、大熊猫和鼠的IL-4核苷酸同源性分别为61.8%、63.4%、69.8%、65.1%、64.1%、64.1%、63.1%、57.0%、60.5%,推导的氨基酸同源性分别为47.2%、50.0%、53.6%、52.1%、48.3%、46.9%、52.1%、43.7%、43.0%。将测序正确的阳性质粒采用脂质体法转染COS-7,用RT-PCR鉴定转染细胞中IL-4基因的表达,并通过MTT法检测到表达的IL-4蛋白具有一定的生物学活性。     

Concentrations of prolactin, luteinizing hormone and follicle stimulating hormone in pituitary and serum of horses: effect of sex, season and reproductive state

Pituitary and serum from 86 male or female horses of various reproductive states were collected in the normal breeding season (summer) and in the nonbreeding season (winter) at a commercial slaughterhouse. Concentrations of prolactin (PRL), luteinizing hormone (LH) and follicle stimulating hormone (FSH) were measured by radioimmunoassay. Concentrations of pregnant mare serum gonadotropin and reproductive steroids in serum and gross appearance of the reproductive tract and gonads were used to catagorize reproductive state. Concentrations of PRL were higher (P less than .01) in summer than in winter in pituitary and serum of mares, stallions and geldings. In summer, mares had higher (P less than .01) concentrations of PRL in serum than stallions. In mares, concentrations of LH in pituitary were higher (P less than .05) in summer than in winter. Concentrations of LH in serum were higher (P less than .01) in summer than in winter in mares and geldings, higher (P less than .01) in mares than in stallions in summer, higher (P less than .01) in geldings than in stallions in summer and higher (P less than .01) in mares with low serum progesterone (P) concentrations than in mares with high P concentrations in summer. Concentrations of FSH in pituitary and serum did not differ between summer and winter for any type of horse.(ABSTRACT TRUNCATED AT 250 WORDS)     

Pituitary hormone and insulin responses to infusion of amino acids and N-methyl-D,L-aspartate in horses

Thirty-nine adult light horse mares, geldings, and stallions were used in two experiments to assess the pituitary hormone and insulin responses to infusions of arginine, aspartic acid, lysine, glutamic acid, and N-methyl-D,L-aspartate (NMA). In Exp. 1, 27 horses were assigned to one of three infusion treatments: 1) physiological saline (1 L); 2) 2.855 mmol of arginine/kg BW in 1 L of water; or 3) 2.855 mmol of aspartic acid/kg BW in 1 L of water. In Exp. 2, 12 horses were assigned, in a multiple-square 4 x 4 Latin square design, to one of four infusion treatments: 1) 2 mL of saline/kg BW; 2) 2.855 mmol of lysine/kg BW in water; 3) 2.855 mmol of glutamic acid/kg BW in water; or 4) 1 mg of NMA/kg BW in water. In Exp. 1, an acute (within 20 min) release of growth hormone (GH) was induced (P = 0.002) by aspartic acid. In contrast, acute release of prolactin (P = 0.001) and insulin (P = 0.002) was induced only by arginine; moreover, the arginine effect on insulin was present only in mares (P = 0.011). In Exp. 2, an acute release of GH was induced (P = 0.001) by glutamic acid and NMA. In males, the glutamic acid-induced GH release was greater than that of NMA; in mares, the NMA-induced GH release was greater than that of glutamic acid (P = 0.069). Both lysine and glutamic acid induced (P = 0.001) acute release of prolactin, whereas an acute release of insulin was elicited (P = 0.002) only by lysine. The NMA-induced LH response was due almost entirely to the response in mares and stallions (P = 0.016), and the NMA-induced FSH release was due almost entirely to the response in mares (reproductive status effect; P = 0.004). In the horse, aspartic acid, glutamic acid, and NMA seem to stimulate GH release; arginine and lysine seem to stimulate prolactin and insulin release; and NMA seems to stimulate LH and FSH release. It seems that N-methyl-D-aspartate glutamate receptors are involved in controlling GH, LH, and FSH secretion, whereas other mechanisms are involved with prolactin secretion. These results also indicate that gonadal steroids interact with amino acid-induced pituitary hormone release in adult horses.     

Comparison of the fractionation and assay of domestic duck and fowl pituitary gonadotrophins

1. Pituitary glycoproteins from domestic ducks and fowls were fractionated to separate luteinising hormone (LH) and follicle stimulating hormone (FSH) activities using the same chromatographic steps. 2. Fractions were bioassayed for LH using the release of progesterone from fowl granulosa cells and for thyroid stimulating hormone (TSH) by measuring the release of thyroxine in 3-d-old chicks. Follicle stimulating hormone activity was measured either in a cockerel-testes radioreceptor assay or by the release of oestrogen from cultured rat Sertoli cells. 3. Fractions containing predominantly FSH or LH activity were isolated from the fowl glycoproteins. Duck gonadotrophin did not occur in fractions corresponding to those containing fowl FSH. 4. Duck gonadotrophin was found in a fraction corresponding with the most highly purified fowl LH fraction. A duck LH fraction was found with little FSH activity for which there was no corresponding fowl LH fraction. 5. It is concluded that domestic fowl and duck gonadotrophins have different chromatographic properties. Further study is required to determine whether the purified duck gonadotrophin preparation comprises proteins with similar physico-chemical properties but with separate FSH and LH biological activities.     

Pubertal changes in the secretion of gonadotropic hormones, testicular gonadotropic receptors and testicular function in the ram

Developmental changes in pituitary content and secretory patterns of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and prolactin (PRL), testicular size and steroidogenic function, testicular LH- and FSH-binding activity, and growth of the accessory sex organs were examined for 24 Dorset X Leicester X Suffolk rams (born in March) every 30 days from 30 to 150 days of age, and again at 200 days. Pituitary LH and FSH contents increased between 30 and 60 days of age and remained constant until 150 days, when contents were somewhat greater than on either 120 or 200 days. LH-pulse amplitude and frequency, and mean FSH concentration, were highest at 60 and (or) 90 days of age. Testicular growth increased dramatically between 90 and 150 days of age in association with increases in the number of LH- (100-fold) and FSH- (33-fold) binding sites in the testis and a small increase in blood testosterone concentration (1 ng/ml). During the same period, pituitary content and blood concentration of PRL increased to maximal values, epididymal, vesicular gland and bulbourethral gland weights increased 6-fold, and body weight doubled. Between 150 and 200 days of age, testosterone concentration increased considerably (8 ng/ml), as did LH-pulse frequency and the amount of LH- and FSH- binding in the testis; the reproductive organs continued to grow at a rate faster than that of the body as a whole. Testicular development of ram lambs was accompanied by increases in the secretion of all three pituitary hormones with gonadotropic properties, and in the number of LH and FSH receptors.     

Fractionation and assay of chicken pituitary hormones

The glycoprotein hormones of the chicken pituitary gland, follicle‐stimulating hormone (FSH), luteinising hormone (LH) and thyroid stimulating hormone (TSH), have been fractionated and partially purified. Fractions were assayed for gonadotrophins using the testicular uptake of ³²P in 1‐d‐old chicks and for thyro‐trophin by chick thyroidal ³²P uptake.     

Reactivity of eleven anti-human leucocyte monoclonal antibodies with lymphocytes from several domestic animals

Nine commercially available monoclonal antibodies and two monoclonal antibodies from The American Type Culture Collection, raised against various human leucocyte surface antigens, were tested on lymphocytes from cow, sheep, goat, swine, horse, cat, dog, mink, and rabbit as well as man. Four antibodies bound to lymphocytes from some of the animals. These were the antibodies against CD8 and CD4 antigen, the antibody to C3b-receptor, and the antibody to the HLA-DR antigen. The CD8 antigen-reactive antibody reacted with lymphocytes from mink, cat, dog, and sheep, while the CD4 antigen-reactive antibody reacted with lymphocytes from mink. The anti-C3b-R antibody reacted with lymphocytes from horse, swine, dog, and cat, and the anti-HLA-DR reacted with lymphocytes from cow, goat, sheep, horse, dog, cat, and mink.     

妊娠期山羊腺垂体中催乳素促卵泡素和促黄体素的分布特点

为了阐明妊娠期垂体中催乳素(PRL)、促卵泡素(FSH),促黄体素(LH)的分布和三者之间协调关系.本试验采用免疫组化SP法检测了PRL、FSH,LH三种激素在妊娠各期山羊的腺垂体细胞中免疫反应产物的分布特点.结果显示,在山羊妊娠过程中,腺垂体中PRL阳性细胞多存在于远侧部,数量逐渐增多,胞质呈强阳性;FSH和LH阳性细胞分布在远侧部和结节部,FSH胞质内颗粒着色深浅不一,LH多核膜着色;相对表达量都逐渐升高,而升高程度不同,表现为妊娠后期LH表达量急剧升高,FSH则变化较小,除了FSH的相对表达量在妊娠前期和中期之间无显著差异(P>0.05)外,3种激素的相对表达量在妊娠前期、中期和后期3个时期之间都存在显著差异(P<0.05),妊娠前、中期FSH的相对表达量均高于PRL和LH,妊娠后期LH的表达量最高.FSH次之,PRL最低.提示PRL,FSH和LH三种激素相互协同以促进胚胎着床并维持妊娠,而且在妊娠早期PRL起主要作用,之后协同FSH和LH维持妊娠.     

Investigations on the Re-establishment of the Positive Feedback of Oestradiol during Anoestrus in the Bitch

To test for the re‐establishment of the positive feedback of oestradiol (E2) during anoestrus in the dog, the hypothalamo–pituitary–ovarian axis of five beagle bitches was challenged by treatments with oestradiol benzoate (EB), mimicking the course of the pro‐oestric E2 secretion. Treatments in anoestrus started 7 days following the decline of progesterone (P) <1 ng/ml; they were repeated in 5 week intervals until onset of pro‐oestrus; another treatment was performed during dioestrus 50 days after onset of the preceding pro‐oestric bleeding. Each dog served as its own control by receiving vehicle‐treatments in one of the following cycles. Each observation period covered a time window of 168 h and blood samples were collected for the determination of luteinizing hormone (LH), follicle‐stimulating hormone (FSH) and E2 in 6 (0–24 h) and 8 h (24–168 h) intervals. In the control periods and as indicated by the parameters area under curve (AUC), basal and maximal values, the availability of LH, FSH and E2 decreased from dioestrus to early anoestrus to increase again during the course of anoestrus (p < 0.05), indicating a gradual desensitization of the hypothalamus towards the negative feedback of oestradiol. At all times treatments with EB lowered the availability of FSH (decreased AUC and basal levels). A delay in the occurrence of the first LH peak after treatments with EB (p < 0.001) and decreased maximal values (p < 0.001) indicated a suppression of the LH‐release. In no case treatment with EB led to a pre‐ovulatory like LH‐surge. In each dog the last trial with EB in anoestrus passed over into pro‐oestrus/oestrus, with a reduced AUC and peak value of the pre‐ovulatory LH‐surge being the only differences to the control group. The observed differences in the response of LH and FSH to treatments with EB point towards subtle differences in the mechanisms controlling the release of these two hormones during anoestrus. From the data obtained, it may be concluded that the time window for E2 to act via a positive feedback seems to be very small and restricted to the end of anoestrus, and that full follicular function is a pre‐requisite to allow for this phenomenon.