ABSTRACT
Aromatase deficiency (AD) is a rare autosomal recessive genetic disease caused by loss-of-function mutations in aromatase gene (CYP19A1), leading to congenital estrogen deficiency syndrome. Both mothers of AD patients during pregnancy and female AD fetus show virilization, while male patients are usually diagnosed in adulthood due to continued height increase and metabolic abnormalities. In 2019, a patient with AD was admitted in the Second Xiangya Hospital. The patient was a 37-year-old adult male who continued to grow linearly after adulthood. His estradiol was below the measurable line, the follicle-stimulating hormone (FSH) increased, bone age delayed, epiphysis unfused, and the bone mass reduced. CYP19A1 gene detection showed that c.1093C>T, p.R365W was homozygous mutation. This disease is rare in clinic. Clinicians need to raise awareness of the disease for early diagnosis and treatment to improve the long-term prognosis of patients.
Subject(s)
Adult , Female , Humans , Male , Pregnancy , 46, XX Disorders of Sex Development/genetics , Aromatase/metabolism , Gynecomastia/genetics , Infertility, Male , Metabolism, Inborn Errors , MutationABSTRACT
At present, there is no reliable in vitro assembled prepubertal testis-like biomimetic organ culture system designed to assess the functional effects of human gonadotropins on Sertoli and Leydig cells. Spermatogenesis is regulated by endocrine, paracrine, and juxtacrine factors (testicular cross-talk), mainly orchestrated by gonadotropins such as luteinizing hormone (LH) and follicle-stimulating hormone (FSH) that play a pivotal role by stimulating Leydig and Sertoli cells, respectively. The aim of our study was to set up an in vitro prepubertal porcine bioengineered construct as a new model for experimental studies on reassembled Sertoli and Leydig cells. We have evaluated Sertoli and Leydig cells obtained from 15- to 20-day-old neonatal pig testes in terms of purity and function. Subsequently, purified Sertoli and enriched Leydig cells were subjected to coincubation to obtain an in vitro prepubertal porcine testis-like culture system. We performed enzyme-linked immunosorbent assay (ELISA) for anti-Müllerian hormone (AMH), inhibin B, and testosterone secretion in the medium, and Real-Time PCR analysis of AMH, inhibin B, FSH-r, aromatase, LHr, and 3β-HSD mRNA expression levels. This in vitro testis-like system was highly responsive to the effects of human gonadotropins and testosterone. AMH mRNA expression and secretion declined, and inhibin-B increased, while FSH-receptor expression was downregulated upon FSH/LH exposure/treatment. Finally, the production of testosterone was increased selectively upon LH treatment. In summary, our proposed model could help to better determine the action of human gonadotropins on Sertoli and Leydig cells. The potential usefulness of the system for shedding light into male infertility-related issues is evident.
Subject(s)
Animals , Male , 3-Hydroxysteroid Dehydrogenases/metabolism , Animals, Newborn , Anti-Mullerian Hormone/metabolism , Aromatase/metabolism , Cell Culture Techniques , Enzyme-Linked Immunosorbent Assay , Follicle Stimulating Hormone/pharmacology , Hormones/pharmacology , In Vitro Techniques , Inhibins/metabolism , Leydig Cells/metabolism , Luteinizing Hormone/pharmacology , Models, Biological , Real-Time Polymerase Chain Reaction , Receptors, FSH/metabolism , Receptors, LH/metabolism , Sertoli Cells/metabolism , Swine , Testis/metabolism , Testosterone/metabolismABSTRACT
Abstract The objective of this study was to evaluate the frequency of C242T polymorphism on the aromatase gene and the allelic and genotypic frequency of these variants in sheep belonging to four breed groups. Blood samples were collected from 187 animals of four breed groups: Dorper, Santa Inês, Texel and White Dorper, originated from herds in the region of Maringá/PR, Brazil. The genomic DNA was extracted using alkaline extraction, with subsequent amplification of the fragments via PCR with specific primer. The samples resulting from amplification were subjected to digestion process using the Dpn II restriction enzyme and to polyacrylamide gel electrophoresis 10.0% and stained with silver nitrate. Three distinct genotypes were observed: homozygous (CC), heterozygous (CT) and homozygous for no cut (TT). The resulting data were analyzed using the POPGENE software with 5% significance. Genotypic frequencies among the breed groups were: Texel (CC - 0.426; CT - 0.511; TT - 0.064), Dorper (CC - 0.073; CT - 0.732; TT - 0.439), White Dorper (CC - 0.021; CT - 0.255; TT - 0.723) and Santa Inês (CC - 0.115; CT - 0.462; TT - 0.423).
Resumo O objetivo deste trabalho foi avaliar as frequências alélicas e genotípicas do polimorfismo do C242T no gene da aromatase em ovinos pertencentes a quatro grupos raciais. Foram coletadas amostras de sangue de 187 animais de quatro grupos raciais: Dorper, Santa Inês, Texel e White Dorper, provenientes de rebanhos da região de Maringá, PR - BR. O DNA genômico foi extraído utilizando o método de extração alcalina, com posterior amplificação dos fragmentos via PCR com primer específico. As amostras resultantes da amplificação foram submetidas ao processo de digestão com auxilio da enzima restrição Dpn II e submetido à eletroforese em gel de poliacrilamida de 10,0% e corado nitrato de prata. Foram observados três genótipos distintos: Homozigoto (CC), heterozigoto (CT) e homozigoto para não corte (TT). Os dados resultantes foram analisados utilizando o software POPGENE com significância de 5%. As frequências genotípicas entre os grupos raciais foram: Texel (CC - 0,426; CT - 0,511; TT - 0,064), Dorper (CC - 0,073; CT - 0,732; TT - 0,439), White Dorper (CC - 0,021; CT - 0,255; TT - 0,723) e Santa Inês (CC - 0,115; CT - 0,462; TT - 0,423).
Subject(s)
Animals , Aromatase/genetics , Gene Frequency , Polymorphism, Genetic , Sheep/genetics , Aromatase/metabolism , Genotype , Sheep/metabolismABSTRACT
The bidirectional communication between oocytes and granulosa cells are mediated by several factors via a local feedback loop(s). The current model was carried out to study the spatial mutual interaction of porcine denuded oocytes and granulosa cells either in direct contact (juxtacrine) or paracrine co-culture using transwell system. Transwell 0.4 µm polyester membrane inserts were used to permit oocytes-granulosa cells paracrine communication with a distance of 2 mm between them in co-culture. Oocytes were cultured with granulosa cells in a defined basic maturation medium for 44 h. In results, oocyte secreted factors (OSFs; GDF9 and BMP15) temporal expression showed progressive decrement by the end of culture in case of direct contact with granulosa cells while it was increased progressively in the paracrine co-culture groups. However, oocytes that were cultured in direct contact showed a significant increase in blastocyst development after parthenogenetic activation than the paracrine co-cultured ones (20% vs. 11.5%, respectively). By the end of culture, granulosa cell count in direct contact showed a significant decrease than the indirect co-culture group (1.2 × 105 cell/mL vs. 2.1 × 105 cell/mL, respectively). Steroids (P4 and E2) and steriodogenesis enzymes mRNA levels showed significant temporal alterations either after 22 h and 44 h of IVM in both juxtacrine and paracrine co-culture systems (P ≤ 0.05). CX43 was much more highly expressed in the granulosa of the direct contact group than the indirect co-culture group. These results indicate the difference in mutual communication between oocytes and granulosa cells that were cocultured either in direct contact (juxtacrine) or with a short distance (paracrine) and propose a new paradigm to study different ovarian follicular cells interaction.
Subject(s)
/genetics , /metabolism , Animals , Aromatase/genetics , Aromatase/metabolism , Bone Morphogenetic Protein 15/genetics , Bone Morphogenetic Protein 15/metabolism , Cell Communication , Cells, Cultured , Coculture Techniques/methods , Connexin 43/genetics , Connexin 43/metabolism , Estradiol/metabolism , Female , Gap Junctions/metabolism , Gene Expression , Granulosa Cells/cytology , Granulosa Cells/metabolism , Growth Differentiation Factor 9/genetics , Growth Differentiation Factor 9/metabolism , Oocytes/cytology , Oocytes/metabolism , Paracrine Communication , Progesterone/metabolism , Reverse Transcriptase Polymerase Chain Reaction , SwineABSTRACT
BACKGROUND: Endometriosis, pro-inflammatory and invasive benign disease estrogen dependent, abnormally express in endometria the enzyme P450Arom, positively regulated by steroid factor-1 (SF-1). Our objective was to study the nuclear protein contents of upstream stimulating factor 2 (USF2a and USF2b), a positive regulator of SF-1, throughout the menstrual cycle in eutopic endometria from women with and without (control) endometriosis and the involvement of nuclear estrogen receptors (ER) and G-coupled protein estrogen receptor (GPER)-1. RESULTS: Upstream stimulating factor 2 protein contents were higher in mid (USF2b) and late (USF2a and USF2b) secretory phase in eutopic endometria from endometriosis than control (p < 0.05). In isolated control epithelial cells incubated with E2 and PGE2, to resemble the endometriosis condition, the data showed: (a) significant increase of USF2a and USF2b nuclear protein contents when treated with E2, PPT (specific agonist for ERa) or G1 (specific agonist for GPER1); (b) no increase in USF2 binding to SF-1 E-Box/DNA consensus sequence in E2-treated cells; (c) USF2 variants protein contents were not modified by PGE2; (d) SF-1 nuclear protein content was significantly higher than basal when treated with PGE2, E2 or G1, stimulation unaffected by ICI (nuclear ER antagonist); and (e) increased (p < 0.05) cytosolic protein contents of P450Arom when treated with PGE2, E2, PPT or G1 compared to basal, effect that was additive with E2 + PGE2 together. Nevertheless, in endometriosis cells, the high USF2, SF-1 and P450Arom protein contents in basal condition were unmodified. CONCLUSION: These data strongly suggest that USF2 variants and P450Arom are regulated by E2 through ERa and GPER1, whereas SF-1 through GPER1, visualized by the response of the cells obtained from control endometria, being unaffected the endogenously stimulated cells from endometriosis origin. The lack of E2 stimulation on USF2/SF-1 E-Box/DNA-sequence binding and the absence of PGE2 effect on USF2 variants opposite to the strong induction that they exert on SF1 and P450 proteins suggest different mechanisms and indirect regulations. The sustained USF2 variants protein expression during the secretory phase in eutopic endometria from women with endometriosis may participate in the pathophysiology of this disease strongly associated with infertility and its characteristic endometrial invasion to ectopic sites in the pelvic cavity.
Subject(s)
Humans , Female , Adult , Aromatase/metabolism , Gene Expression/genetics , Endometriosis/metabolism , Endometrium/metabolism , Estradiol/metabolism , Biopsy , Immunoblotting , Statistics, Nonparametric , Endometriosis/physiopathology , Endometriosis/pathology , Endometrium/cytology , Epithelial Cells/metabolism , Primary Cell Culture , Menstrual Cycle/metabolismABSTRACT
Seasonal variations in the aromatase activity in H. fossilis estimated by a microassay were correlated with the sex steroids, vitellogenin in and ovarian weight during circannual reproductive cycle. In the female catfish, aromatase activity was detectable in the hypothalamus throughout the year whereas in ovary only during active vitellogenesis. In the catfish, hypothalamic aromatase levels increased two times during annual gonadal cycle, once in a fully gravid fish and then in a reproductively quiescent fish. On the other hand, increase in the ovarian aromatase activity was observed only during vitellogenesis, which showed a direct correlation with plasma levels of sex steroids. Further, plasma levels of testosterone and estradiol suggested a precursor-product relationship. At the completion of vitellogenesis, ovarian aromatase activity declined sharply resulting in elevation of plasma testosterone levels, which in turn could be utilized as substrate by the hypothalamic aromatase whose activity was the highest in the postvitellogenic catfish. At least two isoforms of gene, cyp19a and cyp19b, coding for aromatase in ovary and brain respectively were expressed in the catfish. Aromatase activity was more concentrated in those areas of catfish brain, which have been implicated in the control of reproduction.
Subject(s)
Animals , Aromatase/genetics , Aromatase/metabolism , Brain/enzymology , Brain/metabolism , Catfishes/physiology , Circadian Rhythm/physiology , Female , Ovary/enzymology , Ovary/metabolism , Seasons , Substrate SpecificityABSTRACT
OBJECTIVE: This study intends to verify the expression levels and correlation of aromatase, matrix metalloproteinase 2 (MMP-2), matrix metalloproteinase 9 (MMP-9) and CD44 in ductal carcinoma in situ (DCIS) and infiltrating ductal carcinoma (IDC) when both are found in the same breast. METHODS: One hundred and ten cases were evaluated by tissue microarray (TMA) and immunohistochemically screened with anti-aromatase polyclonal antibodies, anti-MMP-2 monoclonal antibodies, anti-MMP-9 policlonal antibodies and anti-CD44 monoclonal antibodies. RESULTS: Aromatase was expressed in IDC and DCIS in 63 (57.3 percent) and 60 (67 percent) of the cases respectively; MMP-2 was similarly expressed in IDC and DCIS in 15 (13.60 percent) cases; MMP-9 was positively expressed in IDC and DCIS in 83 (75.50 percent) and 82 (74.50 percent) cases, respectively; CD44 was positively expressed in IDC and DCIS in 49 (44.50 percent) and 48 (42.60 percent) of the cases, respectively; all of them were highly correlated (p<0,001). The correlation analysis found positive, statistically significant correlation, in IDC between aromatase and MMP-2 (p<0.001) and between aromatase and MMP-9 (p=0.034). Positive correlation between aromatase and MMP-2 (p<0.001) and between MMP-9 and CD44 (p=0.030) were found in DCIS. CONCLUSION: These results allow us to conclude that aromatase through local estrogen synthesis in breast tissue plays an important role in breast carcinogenesis, mainly influencing MMP-2 and MMP-9 which are important participants in tumor cell invasion and dependence of their connection to CD44 for action.
OBJETIVO: O objetivo desse estudo é verificar as expressões e correlações da aromatase, metalloproteinase 2 da matriz (MMP2), metalloproteinase 9 da matriz (MMP-9) e CD44 no carcinoma ductal in situ (CDIS) e carcinoma ductal infiltrativo (CDI) quando ambos estão presentes simultaneamente na mesma mama. MÉTODOS: Foram avaliados 110 casos pelo método de tissue microarray (TMA) e através da utilização de anticorpos policlonais antiaromatase, anticorpos monoclonais anti-MMP-2, anticorpos policlonais anti-MMP-9 e anticorpos monoclonais anti-CD44. RESULTADOS: A aromatase estava expressa de forma positiva no CDI e CDIS em 63 (57,3 por cento) e 60 (67 por cento) casos, respectivamente. A expressão de MMP-2 estava expressa de forma positiva em 15 (13,6 por cento) casos tanto no CDI, quanto no CDIS. A expressão da MMP-9 estava expressa de forma positiva em 83 (75,5 por cento) e 82 (74,5 por cento) casos de CDI e CDIS, respectivamente. A expressão de CD44 estava expressa de forma positiva em 49 (44,5 por cento) e 48 (42,6 por cento) casos de CDI e CDIS, respectivamente. Todos eles apresentando alta correlação (p<0,001). Na avaliação de correlação foi encontrada correlação positiva estatisticamente significante no CDI entre aromatase e MMP-2 (p<0,01) e entre aromatase e MMP-9 (p=0,034). Nos casos de CDIS houve correlação positiva estatisticamente significante entre aromatase e MMP-2 (p<0,001) e entre CD44 e MMP-9 (p=0,030). CONCLUSÃO: Após analisarmos os resultados de nosso estudo, podemos concluir que a aromatase, através da síntese de estrogênio local no tecido mamário, desempenha importante papel na carcinogênese mamária, principalmente influenciando a atuação da MMP-2 e da MMP-9, grandes responsáveis pela invasão celular tumoral que, por sua vez, provavelmente dependem de sua ligação a CD44 para poder desempenhar suas funções.
Subject(s)
Female , Humans , Breast Neoplasms/metabolism , Carcinoma in Situ/metabolism , Carcinoma, Ductal, Breast/metabolism , Neoplasm Proteins/metabolism , /analysis , /metabolism , Aromatase/analysis , Aromatase/metabolism , Immunohistochemistry , /analysis , /metabolism , Matrix Metalloproteinase 9/analysis , Matrix Metalloproteinase 9/metabolismABSTRACT
A deficiência de esteroides gonadais femininos acelera o ganho de massa corpórea, mas os possíveis mecanismos centrais e periféricos envolvidos no aumento da ingestão alimentar e no ganho de massa adiposa que ocorrem nessa condição são pouco conhecidos. Em modelos animais, tanto a falta quanto os defeitos na ação do estrogênio causam aumento da massa corpórea, demonstrando claramente um possível papel desse esteroide no sobrepeso pós-menopausa. Sabe-se que a obesidade e o sobrepeso estão associados a diversas comorbidades que podem levar à morte prematura. Portanto, desvendar os mecanismos relacionados ao ganho de massa corpórea é de grande relevância, assim como desenvolver estratégias que possam prevenir o seu estabelecimento. A regulação do balanço energético está associada ao controle da massa corpórea, sendo o exercício físico um importante modulador desse parâmetro homeostático. Porém, a influência do exercício físico sobre o ganho de massa corpórea durante a deficiência de estrogênio é controversa e depende do protocolo de exercício utilizado. Neste estudo, pretendemos revisar os achados que relacionam a deficiência de estrogênio ao ganho de massa corpórea em animais e seres humanos.
Female steroid hormones deficiency leads to a significant increase in body mass, but the possible central and peripheral mechanisms involved in increased food ingestion and fat accumulation in this situation are still unknown. In animal models, the specific lack of estrogen or its action produce progressive body mass gain, clearly demonstrating the possible role of this hormone in overweight after menopause. Obesity and overweight correspond to a relevant human health problem that can lead to premature death. Therefore unraveling the mechanisms underlying body mass gain is of great relevance, as well as the development of strategies to prevent its establishment. Energy balance regulation is associated with the control of body mass, and physical exercise is an important modulator of this homeostatic parameter. However, the influence of physical exercise in mass gain development during estrogen deficiency is controversial and depends on the exercise protocol used. In this study, we intend to review the data on the effects of estrogen deficiency on body mass gain in humans and animal models.
Subject(s)
Animals , Female , Humans , Mice , Rats , Energy Metabolism/physiology , Estrogens/metabolism , Exercise/physiology , Obesity/metabolism , Aromatase/metabolism , Body Fat Distribution , Body Mass Index , Estrogens/deficiency , Estrogens/genetics , Gene Deletion , Menopause/metabolism , Obesity/genetics , Receptors, Estrogen/genetics , Receptors, Estrogen/metabolismABSTRACT
Background: Endometriosis, a common gynecologic disorder characterized by endometrial glands and stroma outside the uterus, is diagnosed by direct visualization of peritoneal and ovarian implants during laparoscopy. Aim: To study the estrogenic microenvironment in eutopic endometria of women with and without endometriosis. Patients and methods: Eutopic endometria, obtained during laparoscopy from 23 women with endometriosis and 20 fertile cyclic women undergoing tubal sterilization, was studied. P450Arom mRNA expression (RT-PCR) was measured. Also, P450Arom activity was assessed measuring testosterone conversion to estradiol and the concentration of this last hormone in cultured endometrial explants. Results: Age and body mass index was similar in both groups studied. Seventy nine percent of endometria from women with endometriosis and in 29.4 percent from control group expressed P450Arom mRNA (p <0.01). The intensity of the band was higher in secretory endometria from women with endometriosis when compared to controls (p <0.01), but it was similar during the proliferative phase. Estradiol secretion to the culture media by proliferative endometria explants from women with endometriosis was 3-fold higher than secretory endometria (p <0.01) and endometria from control women in both phases. P450Arom activity, in the presence of testosterone, was 7-fold higher in endometrial cultures from women with endometriosis, when compare with the basal culture (p <0.01). However, in endometrial explant cultures from control women, this activity was not statistical different. Conclusions: These results indicate that in women with endometriosis, the microenvironment in the endometria is estrogenic as a consequence of an increased expression and activity of the P450 Arom.
Subject(s)
Female , Humans , Aromatase/metabolism , Endometriosis/metabolism , Endometrium/metabolism , Estrogens/metabolism , Biopsy , Case-Control Studies , Cells, Cultured , Endometriosis/enzymology , Endometriosis/pathology , Endometrium/enzymology , Endometrium/pathology , Estradiol/metabolism , Fertility/physiology , Laparoscopy , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
The mechanisms of high turnover bone loss induced by Cyclosporin A (CsA) are not clearly understood. Deficiencies in sex hormones result in high turnover osteoporosis, and not only androgen but also estrogen plays an important role in maintaining bone mass in men. To study whether or not there are any changes in the levels of sex hormones, aromatization, and the expression of estrogen receptors in CsA-induced osteoporosis, we treated 39 rats with vehicle, low-dose CsA (5 mg/kg) and high dose CsA (15 mg/kg) for 28 days, and measured sex hormone levels by radioimmunoassay. Aromatase activities in ROS cells and 3T3-L1 cells were determined by measuring the conversion rate of 3H-androstenedione into 3H-estrone. ER and ER mRNA were measured by competitive RT-PCR in collected marrow cells and ROS cells. The levels of free testosterone in the serum in low-dose CsA-treated rats were unchanged, but the levels were significantly decreased in those treated with high-dose CsA as previously reported. The levels of total estradiol in the serum were significantly increased in the low-dose CsA-treated group (5 mg/kg) and were comparable to levels of the control group in the high-dose CsA-treated group (15 mg/kg). CsA increased the conversion of 3H-androstenedione to 3H-estrone in ROS cells, but not in 3T3-L1 cells. Meanwhile, CsA treatment did not change the rates of ER or ER mRNA expression in ROS cells or in collected bone marrow cells. In conclusion, CsA treatment decreased the level of free testosterone in the serum, but did not decrease the level of serum estradiol by enhancing aromatization. High-turnover osteoporosis induced by clinical dosage CsA treatment may not be caused by lowering the levels of circulating estrogen or by decreasing the expression of estrogen receptors.
Subject(s)
Male , Mice , Rats , 3T3 Cells , Animals , Aromatase/metabolism , Bone Marrow Cells/metabolism , Cell Line , Cyclosporine/pharmacology , Cyclosporine/adverse effects , Osteoporosis/chemically induced , Rats, Sprague-Dawley , Receptors, Estrogen/metabolism , Gonadal Steroid Hormones/metabolism , Gonadal Steroid Hormones/bloodABSTRACT
The biological action of insulin like growth factor-1 (IGF-1) on follicular steroidogenesis during follicular development in common carp was examined. Studies were carried out by culturing small (1-2 mm diam.) and large (> 2 mm diam.) follicles. IGF-1 (0.3-100 ng/ml) had no effect on progesterone accumulation or aromatase activity during 48 hr culture of small follicles. Progesterone accumulation by large follicles was also unaffected by IGF-1 over the same period, although aromatase activity was stimulated in a dose dependent manner (8-fold increase over basal levels with a maximum stimulatory dose of 30 ng IGF-1/ml). In contrast, small and large follicles responded to IGF-1 in terms of both progesterone accumulation and aromatase activity after longer periods of culture (4 days for progesterone and 6 days for aromatase). Concurrent treatment of small follicles with estradiol (10(-7) M) enhanced the action of IGF-1 on both indices of steroidogenesis and advanced the time at which IGF-1 stimulated activity was first detectable. The effect of estradiol on follicular IGF-1 responsiveness were independent of cell number. In summary, these results demonstrate varied actions of IGF-1 carp ovarian follicular steroidogenesis in vitro. The results indicate that carp follicles acquire responsiveness to IGF-1 in terms of aromatase activity during follicular development in vivo and that estradiol can induce the response in vitro. The results also suggest that estrogen and progesterone biosynthesis by cultured carp ovarian follicles is differentially regulated by IGF-1. Together, these results provide new insights into the biological actions of IGF-1 in fish ovary.
Subject(s)
Animals , Aromatase/metabolism , Culture Techniques , Estradiol/administration & dosage , Estrogens/biosynthesis , Female , Humans , Insulin-Like Growth Factor I/administration & dosage , Ovarian Follicle/drug effects , Progesterone/biosynthesis , Recombinant Proteins/administration & dosage , Steroids/biosynthesisABSTRACT
Oocytes of vitellogenic stage were collected from A. testudineus and incubated in vitro for 4 hr in the absence (control) or presence of 500 ng of piscine gonadotropic hormone (GtH). After the termination of incubation, oocytes were repeatedly washed and then homogenized and ultracentrifuged at 100,000 g to obtain the supernatant fraction (100K sup). Addition of 100K sup from GtH treated oocytes to the oocyte incubation caused a 3-fold increase in ovarian aromatase activity as compared to the control, whereas 100K sup from control oocytes had no such stimulatory activity. Addition of cycloheximide (50 micrograms/ml) along with GtH blocked the stimulatory effect of 100K sup. Treatment of 100K sup from GtH incubate with pepsin or heat also destroyed its stimulatory effect. All these indicate proteinaceous nature of the factor. This factor was purified to 161-fold by utilizing Sephadex G-75 and DEAE Sephacel chromatography. Addition of increasing concentrations of partially purified GtH induced protein (GIP) to oocyte incubation caused a dose dependent increase in ovarian aromatase activity. Both dbcAMP and forskolin mimicked GIP activity. Results indicate that GtH induces the synthesis of a protein factor in perch oocytes which stimulates aromatase activity via the mediation of cAMP.