• The Korean Journal of Zoology
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• v.37 no.3
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• pp.385-408
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• 1994

출생후 0일. 7일. 14일 및 21일 그리고 성체의 흰쥐 전뇌 기저부의 내측중격핵, 수직 및 수평 대각 Broca대 거대세포 시삭전핵 그리고 복부담창구에서 신경성장인자수용체 (nerv-growth 배ctor receptor, NGFr)에 면역반응을 보이는 신경조직과 세포의 분화를 면역조직화학적 및 전자현미경적 방법을 이용하여 조사하였다. 출생후 초기와 성체에서 신경세포 원형질막 뿐만 아니라 세포질에서 NGFr 면역반응이 확인되었다. 그러나 성체에서 신경세포 원형질 막에서의 면역반응은 관찰되지 않았다. 특히 NGFr 면역반응은 골지 부위에서 보였고, 점상의 면역반응물들이 세포체의 세포질과 수상돌기에 소수 분산 분포하였다. 뇌 기저부의 NGFr 면역반응 신경세포들은 뇌 크기의 증대와 뇌 조직의 분화에 따라 점차 수적 증가를 보였다. 이 NGFr 면역반응 신경세포들은 세포의 모양과 세포체의 장 .단축의 비에 따라 6가지 형. 즉 1) 원형. 2) 타원형. 3) 세장형, 4) 방추형, 5) 삼각형, 6) 다각형으로 분류되었다. 전뇌 기저 핵에서 원형과 난형신경세포들의 출현율은 출생후 0일에서 높았으나 성체로 되면서 감소된 반면, 세장형. 방추형, 삼각형 그리고 다각형신경세포들의 출현율은 출생후 0일에서는 낮았으나 성체로 되면서 증가하였다. 모든 핵들에서 NGFr 면역반응 신경세포체의 부피는 출생후 0일에 759-1,640 Um3로 제일 작았으며, 수직 대각 Broca대와 복부담창구에서는 출생후 14일에 각각 5 107 7.385 Um3 그리고 내측중격핵, 수평 대각 Broca대, 거대세포 시삭전핵에서는 출생후 21일에 각각 4,705, 6,061, 6,412 Um3로 최대치를 보였다. 그후 성체로 되면서 모든 핵에서 1,893-3,464 $\mu$m3로 다시 감소하였다. 전자현미경적 관찰에서 출생후 21일된 흰쥐 수평 대각 Broca대에서 NGFr 면역반응은 세포체와 수상돌기의 원형질막 그리고 세포체내에서는 골지체, 다소포성소체 및 조면소포체에서 관찰되었다. 이 결과들로 미루어 NGFr은 출생후 발생단계와 성체의 횐쥐 전뇌 기저부에서 신경세포의 분화와 분포에 관계되는 것으로 생각된다.

• The Korean Journal of Zoology
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• v.36 no.2
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• pp.245-263
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• 1993

신경성장인자 수용체(nerve growth factor receptor, HGFr)의 소재를 휜쥐 전뇌 기저부 핵들의 신경세포와 그 세포내 소기 관에서 연역세포화학적 방법으로 관찰하였다. NGFr에 면역반응을 보이는 신경세포들은 내측중격, 수직 및 수평대각선 브로카대, 거대세포 시삭전핵 그리고 Meynert 기저핵에는 다수 미상핵-피각과 복부담창구에는 소수 관찰 되었다 NGFr에 면역반응을 보이는 신경세포들은 형태학적으로 3가지 형 즉, 1) 난형(또는 원형). 2) 방추형, 3) 삼각형(또는 다각형)으로 구분되었다 내측중격은 주로 난형의 세포로 구성되었으며(91.2%), 수직 및 수평대각선 브로카대, 거대세포 시삭전핵 및 Meynert 기저 핵에는 난형의 세포가 높은 율로 구성되었으나, 방추형과 삼각형 세포들도 내측중격에서보다는 많았다 특히 복부담창구에는 다른 핵들에 비하여 방추형세포(25%)들이 높은 출현율을 보였다 일반적으로 이들 세포의 크기는 삼각형세포가 제일 컸으며, 방추형세포가 그 다음, 그리고 난형 세포가 제일 작았다 전자현미경적 관찰에서 0.05% triton X-100을 처리한 조직중 Meynert 기저핵을 관찰한 결과. Golgi체, multivesicular body 및 소포체들이 N6Fr에 면역반응을 보였으며. trion X-100을 처리하지 않은 조직에서는 단지 수평대각선 브로카대의 신경세포 원형질 막에서만 약한 면역반응을 보였다 위의 결과로 미루어 NGFr은 조연소포체에서 합성되어. Golgi체에서 농축되고, multivesicular body를 통하여 원형질막에 위치하게 되며, 원형질막에서 NGFr은 외래성의 NGF와 복합체를 형성한후, 궁극적으로는 Iysosome의 형태로 세포체 안으로 들어 가는 것으로 추정된다.

• The Korean Journal of Zoology
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• v.36 no.4
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• pp.562-579
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• 1993

출생후 발생에 따른 흰쥐 전뇌 기저부의 Mevnert 기저핵(nucleus basalis of Mevnert)에서 신경성장인자 수용체(nerve growth factor receptor, NGFr)를 함유하는 신경세포의 면역반응성 및 분포, 세포형의 분류, 각 형별 출현율 및 세포 크기 그리고 세포소기관과 NGFr 면역반응과의 관계를 조사하였다. NGFr 면역반응 신경세포들은 출생후 초기 발생에서 세포질 뿐만 아니라 원형질 막에서도 면역반응을 보였으나, 성체에서는 세포질에서만이 면역반응을 보였다. NGFr 면역반응 신경세포는 형태학적 특징인 세포체의 모양과 장·단축의 비를 기준으로 5가지 형, 즉 1) 원형, 2) 난형, 3) 세장형. 4) 방추형, 5) 삼각형(또는 다각형)세포로 구분되었다 원형과 난형 세포는 출생후 0일에서 각각 15.4%. 50.7%의 높은 출현율을 보였으나, 발생이 진행되면서 점차 감소하여 성체에서 각각 7.8%, 28.6%의 출현율을 보였다 반면, 세장형, 방추형 및 삼각형세포는 출생후 0일에서 각각 22%, 2.6% 9.2%의 낮은 출현율을 보였으나, 발생이 진행되면서 지속적으로 증가하여 성체에서는 각각 29 9%, 5.2%, 28.5%의 높은 출현율을 보였다. 출생후 0일에서 신경세포체의 부피는 매우 작았으나(1,642mm3), 발생에 따라 점차 증가하여 14일, 21일에서는 성체에서보다 매우 컸다(각각 6.745, 6,755mm3) 원형 및 난형세포체의 부피는 21일에서(각각 7 955. 7.020mm3), 세장형 방추형 그리고 삼각형세포체의 부피는 14일에서 제일 컸다(각각 7,067, 6,237, 7,748 mm3) 대체적으로 삼각형세포체의 부피가 제일 컸으며, 방추형세포체의 부피가 제일 작았다. 출생후 순일에서의 전자현미경적 관찰에서 세포소기관중 조면소포체, Golgi체, multivesicular body 그리고 세포표면 원형질막이 강한 NGFr 면역반응을 보였다. 위의 결과들로 미루어 MGFr은 출생후 흰쥐 전뇌 기저부 Meynert 기저핵 신경세포의 분화 및 분포와 밀접한 관계를 갖는 것으로 생각된다.

• International Journal of Oral Biology
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• v.37 no.4
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• pp.175-180
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• 2012

Skin-derived precursor cells (SKPs) are multipotent, sphere-forming and embryonic neural crest-related precursor cells that can be isolated from dermis. It is known that the properties of porcine SKPs can be enhanced by leukemia inhibitory factor (LIF) which is an essential factor for the generation of embryonic stem cells in mice. In our present study, to enhance or maintain the properties of murine SKPs, LIF was added to the culture medium. SKPs were treated with 1,000 IU LIF for 72 hours after passage 3. Quantitative real time RT-PCR was then performed to quantify the expression of the pluripotent stem cell specific genes Oct4, Nanog, Klf4 and c-Myc, and the neural crest specific genes Snai2 and Ngfr. The results show that the expression of Oct4 is increased in murine SKPs by LIF treatment whereas the level of Ngfr is decreased under these conditions. Interestingly, LIF treatment reduced Nanog expression which is also important for cell proliferation in adult stem cells and for osteogenic induction in mesenchymal stem cells. These findings implicate LIF in the maintenance of stemness in SKPs through the suppression of lineage differentiation and in part through the control of cell proliferation.

• Molecules and Cells
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• v.40 no.4
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• pp.254-261
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• 2017

Glioblastomas (GBM) are very difficult to treat and their aggressiveness is one of the main reasons for this as well as for the frequent recurrences. MicroRNAs post-transcriptionally regulate their target genes through interaction between their seed sequence and 3'UTR of the target mRNAs. We previously reported that miR-296-3p is regulated by neurofibromatosis 2 (NF2) and enhances the invasiveness of GBM cells via SOCS2/STAT3. In this study, we investigated whether miR-296-5p, which originates from the same precursor miRNA as miR-296-3p, can increase the invasiveness of GBM cells. It was observed that miR-296-5p potentiated the invasion of various GBM cells including LN229, T98G, and U87MG. Through bioinformatics approaches, two genes were identified as miR-296-5p targets: caspase-8 (CASP8) and nerve growth factor receptor (NGFR). From results obtained from Ago2 immunoprecipitation and luciferase assays, we found that miR-296-5p downregulates CASP8 and NGFR through direct interaction between seed sequence of the miRNA and 3'UTR of the target mRNA. Knockdown of CASP8 or NGFR also increased the invasive ability of GBM cells, indicating that CASP8 and NGFR are involved in potentiation of invasiveness by miR-296-5p. Consistent with our findings, CASP8 was downregulated in brain metastatic lung cancer cells, which have a high level of miR-296-5p, compared to parental cells, suggesting that miR-296-5p may be generally associated with the acquisition of invasiveness. Collectively, our results implicate miR-296-5p as a potential cause of invasiveness in cancer and suggest it as a promising therapeutic target for GBM.

• Genomics & Informatics
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• v.16 no.3
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• pp.59-64
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• 2018

Although pork quality traits are important commercially, genome-wide association studies (GWASs) have not well considered Landrace and Yorkshire pigs worldwide. Landrace and Yorkshire pigs are important pork-providing breeds. Although quantitative trait loci of pigs are well-developed, significant genes in GWASs of pigs in Korea must be studied. Through a GWAS using the PLINK program, study of the significant genes in Korean pigs was performed. We conducted a GWAS and surveyed the gene ontology (GO) terms associated with the backfat thickness (BF) trait of these pigs. We included the breed information (Yorkshire and Landrace pigs) as a covariate. The significant genes after false discovery rate (<0.01) correction were AFG1L, SCAI, RIMS1, and SPDEF. The major GO terms for the top 5% of genes were related to neuronal genes, cell morphogenesis and actin cytoskeleton organization. The neuronal genes were previously reported as being associated with backfat thickness. However, the genes in our results were novel, and they included ZNF280D, BAIAP2, LRTM2, GABRA5, PCDH15, HERC1, DTNBP1, SLIT2, TRAPPC9, NGFR, APBB2, RBPJ, and ABL2. These novel genes might have roles in important cellular and physiological functions related to BF accumulation. The genes related to cell morphogenesis were NOX4, MKLN1, ZNF280D, BAIAP2, DNAAF1, LRTM2, PCDH15, NGFR, RBPJ, MYH9, APBB2, DTNBP1, TRIM62, and SLIT2. The genes that belonged to actin cytoskeleton organization were MKLN1, BAIAP2, PCDH15, BCAS3, MYH9, DTNBP1, ABL2, ADD2, and SLIT2.

• IMMUNE NETWORK
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• v.2 no.3
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• pp.133-136
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• 2002

Background: The costimulatory molecule 4-1BB, a member of nerve growth factor receptor/tumor necrosis factor (NGFR/TNFR) super family, is involved in cell survival and death. Methods: In this study, female C57BL/6 ($H-2^b$) mice were used as a recipient, and DBA/2 ($H-2^d$) as a donor to assess a mixed lymphocyte reaction (MLR) and CTL response in vitro, and skin graft survival. IL-2, IFN level was measured by ELISA. Results: Mixed lymphocyte reaction (MLR) analysis showed that 4-1BB-deficient responder cells showed enhanced cellular proliferation over littermate controls. In contrast, IL-2 production was diminished only in 4-1BB knockout cultures. The IFN expression, on the other hand, was comparable between the groups. When female C57BL/6 ($H-2^b$) mice were grafted with the trunk skin of DBA/2 ($H-2^d$) mice, the in vivo tissue destruction of 4-1BB-deficient mice was not distinct from the normal littermates. Conclusion: These data suggest that 4-1BB is critical for the induction of alloreactive responses in vitro but 4-1BB alone could not change the course of skin rejection in vivo.

• International Journal of Oral Biology
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• v.35 no.4
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• pp.209-214
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• 2010

Cells that have endogenous multipotent properties can be used as a starting source for the generation of induced pluripotent cells (iPSC). In addition, small molecules associated with epigenetic reprogramming are also widely used to enhance the multi- or pluripotency of such cells. Skinderived precursor cells (SKPs) are multipotent, sphereforming and embryonic neural crest-related precursor cells. These cells can be isolated from a juvenile or adult mammalian dermis. SKPs are also an efficient starting cell source for reprogramming and the generation of iPSCs because of the high expression levels of Sox2 and Klf4 in these cells as well as their endogenous multipotency. In this study, valproic acid (VPA), a histone deacetylase (HDAC) inhibitor, was tested in the generation of iPSCs as a potential enhancer of the reprogramming potential of SKPs. SKPs were isolated from the back skins of 5-6 week old C57BL/6 X DBA/2 F1 mice. After passage 3, the SKPs was treated with 2 mM of VPA and the quantitative real time RT-PCR was performed to quantify the expression of Oct4 and Klf4 (pluripotency specific genes), and Snai2 and Ngfr (neural crest specific genes). The results show that Oct4 and Klf4 expression was decreased by VPA treatment. However, there were no significant changes in neural crest specific gene expression following VPA treatment. Hence, although VPA is one of the most potent of the HDAC inhibitors, it does not enhance the reprogramming of multipotent skin precursor cells in mice.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.31 no.3
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• pp.239-247
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• 2005

This study demonstrated that xenogenic human marrow mesenchymal stem cells (hMSCs) could elicit the regeneration of the sensory nerve after axotomy in the adult rats infraorbital nerves without immunosuppression. For this, we evaluated the behavioral testing for functional recovery of the nerve and histological findings at weeks 3 and 5 compared to controls. Xenogenic hMSCs did not evoke any significant inflammatory or immunologic reaction after systemic and local administrations. HMSCs-treated rats exhibited significant improvement on sensory recovery tested with von Frey monofilaments. At 5 postoperative weeks, in the hMSCs treated nerve, expression of myelin basic protein (MBP), neurofilament (NF) at the site of axotomy was higher than control. And mRNA expression of neurotropin receptor Trk precursor (TrkPre), nerve growth factor receptor (NGFR) and neuropeptide (NPY) in trigeminal ganglion were also higher. The number of myelinated nerve at distal stump and cells in trigeminal ganglion were higher in hMSC treated rats. So it was supposed that transplanted MSCs contributed to reducing post-traumatic degeneration and production of neurotrophic factors. Immunofluorescence labeling showed small portion of hMSCs (<10%) expressed a phenotypic marker of Schwann cell (S-100). Xenogenic or allogenic mesenchymal stem cells might have immune privileged characteristics and useful tool for cell based nerve repair.

• Journal of Life Science
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• v.19 no.1
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• pp.21-33
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• 2009

In this study, we investigated the effect of Rhei Rhizoma (RR; 大黃) water extract on gene expression in a hypoxia model of cultured rat hippocampal neurons. RR water extract $(2.5{\mu}g/ml)$ was added to the culture media on day 10 in vitro (DIV10), and a hypoxic shock (2% $O_2$/5% $CO_2$, $37^{\circ}C$, 3 h) was given on DIV13. After maintaining the cultures in normoxia for 24 hr, total RNA was isolated and used for microarray analysis. The MA-plot indicated that most genes were up- or downregulated within 2-fold. There were more downregulated genes (725 ea) than upregulated ones (472 ea) when larger than Global M value 0.2 (i.e., >15% increase) or smaller than Global M value -0.2 (i.e., >15% decrease) were considered. Antiapoptosis genes such as Tegt (2.4-fold), Nfkb1 (2.4-fold) Veg (1.8-fold), Ngfr (1.6-fold) were upregulated, while pro-apoptosis genes such as Bad (-64%), Cstb (-66%) were downregulated. Genes for combating environmental stress (stress response genes) such as Defb3 (2.7-fold), Cygb (2.2-fold), Ahsg (2.18-fold), Alox5 (2-fold) were upregulated. Genes for cell proliferation (cell cycle-related genes) such as Erbb2 (1.84-fold), Mapk12 gene (1.8-fold) was upregulated. Therefore, RR water extracts upregulate many pro-survival genes while downregulating many pro-death genes. It is interpreted that these genes, in combination with other regulated genes, can promote neuronal survival in a stress such as hypoxia.