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Kyu Pil Lee,Ju-Hong Jeon,Insuk So,Ki Whan Kim 대한생리학회-대한약리학회 2006 The Korean Journal of Physiology & Pharmacology Vol.10 No.4
Many gastrointestinal muscles show electrical oscillation, so-called "slow wave", originated from interstitial cells of Cajal (ICCs). Thus, a technique to freshly isolate the cells is indispensable to explore the electrophysiological properties of the ICCs. To apply an enzyme solution on the serosal surface for cell isolation, the intestine was inverted and 0.02% trypsin solution and 0.04% collagenase solution were applied to serosal cavity. After the enzyme treatment, mucosal layer was removed and longitudinal muscle layer was gently separated from the rest of tissue. The thin layer was stretched in the recording chamber and mounted on an inverted microscope. Using β-escine, perforated whole cell patch clamp technique was used. Under a microscope, the tissue showed smooth muscle cells and interstitial cells around the myenteric plexus. Under voltage clamp condition, three types of membrane potential were recorded. One group of interstitial cells, which were positive to methylene blue and CD34, showed spontaneous outward current. These cells had bipolar shape and were considered as fibroblast-like cells because of their peculiar shape and arrangement. Another group, positive to c-kit and methylene blue, showed spontaneous inward current. These cells had more rounded shape and processes and were considered as ICCs. The third, positive to c-kit and had granules containing methylene blue, showed quiet membrane potentials under the voltage-clamp mode. These cells appeared to be resident macrophages. Therefore, in the freshly isolated thin tissue preparation, methylene blue could easily identify three types of cells rather than morphological properties. Using this method, we were able to study electrical properties of fibroblast and residential macrophage as well as myenteric ICCs.
Insuk So,이규필,Jae Yeoul Jun,In-Youb Chang,서석효,김기환 한국분자세포생물학회 2005 Molecules and cells Vol.20 No.3
Classical transient receptor potential channels (TRPCs) are thought to be candidates for the nonselective cationchannels (NSCCs) involved in pacemaker activity and its neuromodulation in murine stomach smooth muscle. We aimed to determine the role of TRPC4 in the formation of NSCCs and in the generation of slow waves. At a holding potential of –60 mV, 50 μM carbachol (CCh) induced INSCC of amplitude [500.8 ±161.8 pA (n = 8)] at –60 mV in mouse gastric smooth muscle cells. We investigated the effects of commercially available antibodies to TRPC4 on recombinant TRPC4expressed in HEK cells and CCh-induced NSCCs in gastric smooth muscle cells. TRPC4 currents in HEK cells were reduced from 1525.6 ± 414.4 pA (n = 8) to146.4 ± 83.3 pA (n = 10) by anti-TRPC4 antibody and INSCC amplitudes were reduced from 230.9 ± 36.3 pA (n= 15) to 49.8 ± 11.8 pA (n = 9). Furthermore, INSCC in the gastric smooth muscle cells of TRPC4 knockout mice was only 34.4 ± 10.4 pA (n = 8) at –60 mV. However, slow waves were still present in the knockout mice. Our data suggest that TRPC4 is an essential component of the NSCC activated by muscarinicstimulation in the murine stomach.
Gene transfer of the K<sub>ATP</sub> channel restores age-related erectile dysfunction in rats
So, Insuk,Chae, Mee Ree,Lee, Sung Won BLACKWELL SCIENCE 2007 BJU INTERNATIONAL Vol.100 No.5
<P>OBJECTIVE</P><P>To determine if gene transfer of the ATP-sensitive potassium (K<SUB>ATP</SUB>) channel can reverse age-related erectile dysfunction in the rat, as the K<SUB>ATP</SUB> channel is an important subtype of potassium channels regulating smooth muscle tone.</P><P>MATERIALS AND METHODS</P><P>In an <I>in vitro</I> study, gene were transferred using cDNA of the K<SUB>ATP</SUB> channel in cultured human corporal smooth muscle (CSM) cells and human embryonic kidney (HEK) cells. After gene transfer, the activities of transferred channels were assessed by the patch-clamp technique. In an <I>in vivo</I> study, 15 old rats were used for groups of gene therapy and nine young adult rats were used as normal controls. The old rats were divided into three groups, i.e. controls and two gene-transfer groups (Kir6.1 + SUR2B and Kir6.2 + SUR2B). The intracavernosal pressure (ICP) response to cavernosal nerve stimulation was assessed after intracorporal injection with naked cDNA of the K<SUB>ATP</SUB> channel. The transgene expression of the K<SUB>ATP</SUB> channel was examined by reverse transcription-polymerase chain reaction (RT-PCR) in rats transfected with cDNA of Kir 6.1 and 6.2.</P><P>RESULTS</P><P>The transferred gene of the K<SUB>ATP</SUB> channel was functionally active and appropriate for gene transfer. The mean (<SMALL>SEM</SMALL>) ratio of ICP to systemic blood pressure in the gene-transfer groups, at 79.4 (1)% and 76.5 (2.6)% (both eight rats) was significantly higher than that in age-matched control rats, at 59.4 (3.3)% (eight), and similar to that in the young control rats, at 77.1 (2.7)% (nine). The RT-PCR showed expression of Kir6.1 and 6.2 genes in the transfected groups.</P><P>CONCLUSION</P><P><I>In vivo</I> gene transfer of the K<SUB>ATP</SUB> channel can physiologically restore erectile function in aged rats, and might be applicable to the development of new forms of therapy for treating human erectile dysfunction.</P>
Imaging Phenotyping Using Radiomics to Predict Micropapillary Pattern within Lung Adenocarcinoma
Song, So Hee,Park, Hyunjin,Lee, Geewon,Lee, Ho Yun,Sohn, Insuk,Kim, Hye Seung,Lee, Seung Hak,Jeong, Ji Yun,Kim, Jhingook,Lee, Kyung Soo,Shim, Young Mog Elsevier 2017 Journal of thoracic oncology Vol.12 No.4
The antitumor effects of geraniol: Modulation of cancer hallmark pathways (Review)
CHO, MINSOO,SO, INSUK,CHUN, JUNG NYEO,JEON, JU-HONG D.A. Spandidos 2016 International journal of oncology Vol.48 No.5
<P>Geraniol is a dietary monoterpene alcohol that is found in the essential oils of aromatic plants. To date, experimental evidence supports the therapeutic or preventive effects of geraniol on different types of cancer, such as breast, lung, colon, prostate, pancreatic, and hepatic cancer, and has revealed the mechanistic basis for its pharmacological actions. In addition, geraniol sensitizes tumor cells to commonly used chemotherapy agents. Geraniol controls a variety of signaling molecules and pathways that represent tumor hallmarks; these actions of geraniol constrain the ability of tumor cells to acquire adaptive resistance against anticancer drugs. In the present review, we emphasize that geraniol is a promising compound or chemical moiety for the development of a safe and effective multi-targeted anticancer agent. We summarize the current knowledge of the effects of geraniol on target molecules and pathways in cancer cells. Our review provides novel insight into the challenges and perspectives with regard to geraniol research and to its application in future clinical investigation.</P>
Byung Joo Kim,So Yeon Lee,Hyung Woo Kim,Eun-Jung Park,Jun Kim,Sang Jeong Kim,Insuk So,Ju-Hong Jeon 대한생리학회-대한약리학회 2009 The Korean Journal of Physiology & Pharmacology Vol.13 No.5
Cerebellar Purkinje cells (PCs) play a crucial role in motor functions and their progressive degeneration is closely associated with spinocerebellar ataxias. Although immunohistochemical (IHC) analysis can provide a valuable tool for understanding the pathophysiology of PC disorders, the method validation of IHC analysis with cerebellar tissue specimens is unclear. Here we present an optimized and validated IHC method using antibodies to calbindin D28k, a specific PC marker in the cerebellum. To achieve the desired sensitivity, specificity, and reproducibility, we modified IHC analysis procedures for cerebellar tissues. We found that the sensitivity of staining varies depending on the commercial source of primary antibody. In addition, we showed that a biotin-free signal amplification method using a horseradish peroxidase polymer-conjugated secondary antibody increases both the sensitivity and specificity of ICH analysis. Furthermore, we demonstrated that dye filtration using a 0.22Ռm filter eliminates or minimizes nonspecific staining while preserving the analytical sensitivity. These results suggest that our protocol can be adapted for future investigations aiming to understand the pathophysiology of cerebellar PC disorders and to evaluate the efficacy of therapeutic strategies for treating these diseases.