• 한국발생생물학회지:발생과생식
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• 제26권1호
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• pp.13-21
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• 2022

Neurokinin B (NKB) is a neuropeptide involved in the regulation of reproductive endocrine system of vertebrate animals, including fish. However, the pathway of NKB action in fish has not been clearly elucidated. In order to clarify the effect of NKB and NKF (neurokinin F) on gonadotropic hormone (GTH) gene expression in the pituitary, we studied the changes of LHβ and FSHβ gene expressions by using two different pituitary culture methods (whole pituitary culture or dispersed pituitary cell culture). Pituitaries were removed from mature female and male Nile tilapia. Changes of LHβ and FSHβ gene expressions were measured and compared after the treatment with NKB or NKF peptides at concentrations 0 to 1,000 nM. Expression of GTH genes in the whole pituitary cultures treated with NKB or NKF peptides did not show significant difference except in female at one concentration when treated with NKF. On the contrary, there were significant changes of GTH gene expressions in the dispersed pituitary cell cultures when treated with NKB and NKF peptides. These results suggest that dispersed pituitary cell culture is more relevant than whole pituitary culture in studying the function of pituitary, and that NKB and NKF could act directly on the pituitary to regulate the expression of GTH genes.

• 한국발생생물학회지:발생과생식
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• 제4권2호
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• pp.237-242
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• 2000

흰쥐 시상하부에서 합성ㆍ분비되어 뇌하수체 전엽에서의 growth hormone (GH) 분비를 촉진하는 growth hormone releasing hormone (GHRH)이 시상하부 이외 조직들 (extrahypothalamic tissues)인 태반, 생식소, 그리고 뇌하수체 전엽에서도 발현됨이 보고되었다. 본 연구는 흰쥐 뇌하수체 전엽에서 발현되는 GHRH의 기능을 조사하기 위해 i)세포 배양을 시행하면서 GHRH의 세포내 함량, 분비 그리고 세포분획법 (cell-fractionation)을 사용하여 분리한 뇌하수체 세포 유형별로 GHRH 함량을 방사면역측정법으로 조사하였고, ii)체외배양 중인 뇌하수체 전엽세포의 증식에 미치는 GHRH의 효과를 측정하기 위해 [$^3$H] thymidine incorporation assay를, 그리고 iii) GHRH의 세포분열 촉진 효과와 세포내 c-fos 유전자 발현과의 상관관계를 조사하기 위해 northern blot analysis를 시행하였다. GHRH 방사면역측정법을 시행한 결과 상당량의 GHRH-like 분자들이 흰쥐 뇌하수체 전엽내에 존재하고, 체외 세포배양시 분비됨을 관찰하였다. 세포분획을 사용한 실험에서 GHRH 함량은 gonadotrope, somatotrope, lactotrope 그리고 thyrotrope 순으로 나타났다. 이 러한 결과는 흰쥐 뇌하수체 전엽에서 생성된 GHRH가 국부적인 조절인자, 특히 상이한 유형의 세포들 간의 상호조절 (cross-talk)을 통해 뇌하수체 전엽에서의 세포분열과 분화, 그리고 기능조절에 관여할 가능성을 보여주었다. GHRH는 체외 배양중인 뇌하수체 전엽세포의 [$^3$H] thymidine incorporation을 농도의존적으로 증가시켰으며, 이러한 GHRH의 세포분열 촉진 효과는 예상대로 세포내 oncogene 활성 의 증가를 통해 일어나는 것임을 c-fos northrn blot으로 확인하였다. 결론적으로, 본 연구는 흰쥐 뇌하수체 전엽에서 합성되는 GHRH가 paracrine 또는 autocrine 기작으로 GH의 분비 촉진 이외에도 세포분열의 조절함을 시사하는 것이다.

• Journal of Korean Neurosurgical Society
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• 제54권2호
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• pp.132-135
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• 2013

The small pituitary mass was incidentally found in 40-years-old women with renal cell carcinoma. The endocrinological and ophthalmological evaluation revealed no deficit and the short-term follow-up was recommended. In 6 months later, the visual disturbance was reported and the size of mass was increased. The tumor was removed totally via the trans-sphenoid approach. The post-operative endocrinological insufficiency was not noticed. During one year of follow-up period, there was no evidence of recurrence without adjuvant radiotherapy. The clinical features of pituitary metastasis from renal cell carcinoma were similar to those of pituitary adenoma. The possibility of pituitary metastasis should be kept in mind in patients with sellar mass and renal cell carcinoma.

• 한국발생생물학회지:발생과생식
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• 제2권2호
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• pp.189-196
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• 1998

Growth Hormone Releasing Hormone (GHRH)은 척추동물의 시상하부로부터 합성, 분비되어 시상하부-뇌하수체간의 문맥계를 통해 뇌하수체 전엽에 작용하여 Growth Hormone (GH)의 분비를 촉진한다. 시상하부에서 발현되는 일부 Releasing Hormone 들이 여러 시상하부외 조직에서도 검출되고 조직특이적인 기능을 수행한다는 사실이 여러 연구자들에 의해 밝혀졌다. 이러한 사실들을 배경으로 본 연구자는 GHRH가 흰쥐의 뇌하수체 전엽과 뇌하수체로부터 유래된 종양세포주들에서 발현될 가능성을 조사하였다. GHRH 펩타이드와 mRNA의 존재와 구조를 규명하기 위하여 뇌하수체와 배양 세포를 사용하여 GHRH immunocytochemistry, 방사면역측정법, GHRH PCR과 RNase protection assay를 시행하였다. Immunocytochemistry의 결과 gonadotrope (대형)와 somat-olactotrope (중간형)로 추정되는 세포들에서 GHRH 염색이 나타났고, Somatolactotrope성 종양세포인 GH3 cell 추출물에서 immunoreactive GHRH가 방사면역측정법으로 검출되었다. 3'rapid amplification of cDNA end (3'-RACE)를 시행한 결과, 흰쥐 뇌하수체에 GHRH transcript가 존재하고, 그 3'end 부분이 다른 조직내의 GHRH와 동일함을 확인하였다. GHRH RT-PCR에서도 뇌하수체와 종양세포주들인 $\alpha$T3 cell (gonadotrope성)과 GH3 cell에서 예상 산물들이 증폭되었다. RNase protection assay를 시행한 결과 난소절제에 의해 뇌하수체내 GHRH 유전자 발현이 증가됨을 확인하였다. 이상의 결과는 GHRH가 뇌하수체 전엽의 gonadotrope와 somatotrope에서 발현되고, paracrine 또는 autocrine조절물질로 작용하여 GH 분비 외에도 뇌하수체 전엽 세포들의 분화와 분열등에 관여함을 시사한다.

• The Korean Journal of Physiology and Pharmacology
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• 제23권3호
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• pp.171-179
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• 2019

Pituitary tumors are usually benign but can occasionally exhibit hormonal and proliferative behaviors. Dysregulation of the G1/S restriction point largely contributes to the over-proliferation of pituitary tumor cells. F-box protein S-phase kinase-interacting protein-2 (SKP2) reportedly targets and inhibits the expression of $p27^{Kip1}$, a well-known negative regulator of G1 cell cycle progression. In this study, SKP2 expression was found to be upregulated while $p27^{Kip1}$ expression was determined to be downregulated in rat and human pituitary tumor cells. Furthermore, SKP2 knockdown induced upregulation of $p27^{Kip1}$ and cell growth inhibition in rat and human pituitary tumor cells, while SKP2overexpression elicited opposite effects on $p27^{Kip1}$ expression and cell growth. The expression of microRNA-186 (miR-186) was reported to be reduced in pituitary tumors. Online tools predicted SKP2 to be a direct downstream target of miR-186, which was further confirmed by luciferase reporter gene assays. Moreover, miR-186 could modulate the cell proliferation and $p27^{Kip1}$-mediated cell cycle alternation of rat and human pituitary tumor cells through SKP2. As further confirmation of these findings, miR-186 and $p27^{Kip1}$ expression were downregulated, while SKP2 expression was upregulated in human pituitary tumor tissue samples; thus, SKP2 expression negatively correlated with miR-186 and $p27^{Kip1}$ expression. In contrast, miR-186 expression positively associated with $p27^{Kip1}$ expression. Taken together, we discovered a novel mechanism by which miR-186/SKP2 axis modulates pituitary tumor cell proliferation through $p27^{Kip1}$-mediated cell cycle alternation.

• 한국가금학회지
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• 제23권3호
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• pp.113-119
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• 1996

A series of experiments were conducted to investigate the role of protein kinase C (PKC) as a second messenger in vasoactive intestinal peptide (VIP) mediated prolactin secretion. Primary pituitary cells (106 cells/treatment) were separated from laying hens and incubated in M-199 with 5% chicken serum and 5% fetal calf serum. The VIP(0.1 $\pi$M) treatment enhanced prolactin Secretion into media upto 9-fold during 48-h incubation. The phorbol 12-myristate 13-acetate (PMA), a PKG agonist, increased prolactin secretion upto 2-fold at 0.1 nM PMA (P<0.01), and the prolactin secretion was not significantly higher than this concentration. Staurosporine (ST; 1.0$\pi$M) a PKC antagonist, decreased by 70% of 0.1 $\pi$M VIP-stimulated prolactin secretion and by 48% of 10 ${\mu}$M PMA-stimulated prolactin secretion (P<0.01). However, pituitary cell prolactin content did not differ in any treatment (P>0.05). In conclusion, these results indicate that the PKC second messenger system is involved in VIP-stimulated prolactin release in chicken primary pituitary cell culture.

• Archives of Pharmacal Research
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• 제26권1호
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• pp.34-39
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• 2003

The traditional Asian medicinal herb, roots of Astragalus (A.) membranaceus (Leguminosae), is used for many purposes, some of which are purported to stimulate the release of growth hormone in vivo. Extracts of A. membranaceus were tested to determine whether they stimulate the release of growth hormone in rat pituitary cell culture. A. membranaceus was extracted sequentially with 80% ethanol (fraction A), n-hexane (fraction B); the test compound from the herbal extraction was isolated using silica gel column chromatography and was identified with spectral data. Test compound was also extracted by traditional boiling water methods. Induction of growth hormone in pituitary cell culture was conducted with isolated compounds and extracted fractions of A. Radix (dried roots of A. membranaceus). The fraction A was not active in the rat pituitary cell culture, but the fraction B derived from the ethanol fraction stimulated the release of growth hormone in culture. Six compounds from fraction B (1-6) were isolated and identified previously. The compounds 1,2-benzendicarboxylic acid diisononylester (1), $\beta$-sitosterol (2), and 3-Ο-$\beta$-D-galactopyranosyl-$\beta$-sitosterol (5) did not induce growth hormone release in the culture. Formononetin (3), 9Z, 12Z-octadecadienoic acid (4), stigmast-4-en-6$\beta$-o1-3-one (6) and 98-E, a mixture of 1'-9, 12-octadecadienoic acid (Z,Z)-2',3'-dihydroxy-propylester (7) and 1'-hexadecanoic acid-2',3'-dihydroxy-propylester (8) stimulated the release of growth hormone in the rat pituitary cell culture significantly compared to the control. In conclusions, four compounds isolated from extracts of A. Radix induced growth hormone release in the rat pituitary cell culture. The 98-E isolate was the most active inducer of growth hormone release.

• 생약학회지
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• 제9권1호
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• pp.33-39
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• 1978

The present study is involved with the prolactin secretion from anterior pituitary gland by ginseng saponin since it was handled down by tradition that ginseng might influence the milk secretion when it was given to nursing mother. To investigate the effect of saponin on the prolactin production or release from the anterior pituitary gland, cell culture study and whole animal studies were carried out. For the cell culture study, enzymatically dispersed anterior pituitary cells of rat anterior pituitary gland in HEPES buffers containing trypsin were used. Ginseng saponin was added to the culture media and the amount of prolactin produced in the cell culture media was determined by radloimmunoassay(RIA) technique. Dose-dependent increases of prolactin with ginseng saponin were observed, whereas, no change was observed without ginseng treatment. For the whole animal study, normal and castrated rats which previously cannulated into the heart via the right juglar vein were used. The prolactin concentration in plasma were determined by using the technique of RIA. In normal rats, prolactin concentration in plasma were elevated dramatically after 1 hour of ginseng saponin administration, whereas, instantaneous increases were observed in castrated rats. For prolactin assay by RIA, NIAMDD Rat Prolactin Kit and NIAMDD Rat Prolactin RP-1 were used as standard. The results indicate that ginseng saponins increase the release of prolactin from the anterior pituitary gland and production of prolactin from the cell in rats.

• 대한의생명과학회지
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• 제19권4호
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• pp.295-302
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• 2013

Some of the pituitary adenomas are invasive and spread into neighboring tissues. In previous studies, the invasion of pituitary adenomas is thought to be associated with epithelial-mesenchymal transition (EMT). In addition to that, we thought that mesenchymal stem cells (MSCs) exist in relevant microenvironment in pituitary adenoma. However, it has been little known about the existence of MSCs from pituitary adenoma. So we investigated whether mesenchymal stem-like cells (MSLCs) can be isolated from the pituitary adenoma specimen. We isolated and cultured candidate MSLCs from the fresh pituitary adenoma specimen with the same protocols used in culturing bone marrow derived MSCs (BM-MSCs). The cultured candidate MSLCs were analyzed by fluorescence-activated cell sorting (FACS) for surface markers associated with MSCs. Candidate MSLCs were exposed to mesenchymal differentiation conditions to determine the mesenchymal differentiation potential of these cells. To evaluate the tumorigenesis of candidate MSLCs from pituitary adenoma, we implanted these cells into the brain of athymic nude mice. We isolated cells resembling BM-MSCs named pituitary adenoma stroma mesenchymal stem-like cells (PAS-MSLCs). PAS-MSLCs were spindle shaped and had adherent characteristics. FACS analysis identified that the PAS-MSLCs had a bit similar surface markers to BM-MSCs. Isolated cells expressed surface antigen, positive for CD105, CD75, and negative for CD45, NG2, and CD90. We found that these cells were capable of differentiation into adipocytes, osteocytes and chondrocytes. Tumor was not developed in the nude mice brains that were implanted with the PAS-MSLCs. In this study, we showed that MSLCs can be isolated from a pituitary adenoma specimen which is not tumorigenic.

• Archives of Pharmacal Research
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• 제3권2호
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• pp.85-86
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• 1980

Dopaminergic influences on prolactin release from lactotrophs have been studied using the rat anterior pituitary cell culture. The prolactin inhibiting activity of hypothalamic extracts was examined in relation to dopamine. Dopamine inhibited prolactin secretion from the rat anterior pituitary cell culture in a dose dependent fashion. The median effective dose was $2{\times}10^{-7}$ / M and the maximal inhibition (70-90 % of the control value) was shown by 10$^{-5}$ / M dopamine. Further increase in dopamine concentration did not result in any further inhibition of prolactin secretion.nhibition of prolactin secretion.