Chromogenic In Situ Hybridisation (CISH): Experiences from Histomorphological Research on Cleft Lip and Palate Tissue Samples

Activity: Talk or presentation types › Oral presentation

Description

Chromogenic In-situ Hybridization (CISH) is a relatively new technique that allows the localization and visualization of a specific DNA or RNA sequence in a tissue specimen. The technique utilizes a chromogen-labelled DNA probe that gives a chromogenic dot-like response that can be visualized using the peroxidase reaction. Since the CISH probes are standardized with complete kits, the need to perform the more expensive fluorescent microscopy is eliminated. Given this superiority of the technique, we aimed to explore the utility of the technique in cleft lip and palate tissue samples. Cleft lip and palate are one of the most common congenital defects worldwide. The exact morpho-etiopathogenesis that cause and sustain the impaired wound healing in cleft lip and palate are yet to be elucidated, making the application of CISH extremely beneficial in morphological research.. CISH was performed using ZytoDot 2C CISH Implementation Kit (ZytoVision GmbH, Bremerhaven, Germany). Probes of various growth factors like FGFR1, FGFR2, and FOXO1 were used in this study. Additional probes of homeobox genes like DLX4, HOXB3, MSX2, and PTX3 were also used. The study comprised samples from 15 pediatric patients suffering from cleft lip and palate. Under the microscope, two brown-colored dots (signals) were expected per nuclei of normal cells in interphase or metaphase without aberrations of the examined chromosomes.. No changes in the gene amplification and expression of homeobox genes like DLX4, HOXB3, MSX2, and PTX3 were noted in the cleft-affected tissue. Similar results were noted for FGFR1, FOXO1, and FGFR2. In the connective tissue, however, FGFR1 demonstrated a low level of amplification, suggesting a downstream dysregulation in wound healing pathways.. Due to the easy interpretation of results and the technique’s superiority when compared with other morphological techniques, CISH represents an emerging modality in morphological tissue research.
Period29 Mar 2023
Event titleRSU International Research Conference 2023: Knowledge for Use in Practice
Event typeConference
OrganiserRīga Stradiņš University
LocationRiga, LatviaShow on map
Degree of RecognitionInternational

Related content