The expression of Pannexin 1 in the developing human spinal cord and dorsal root ganglia

Objectives: The objective of this study was to explore the expression of Pannexin 1 (Panx1) in the spinal cord (SC) and ganglia of human conceptus during early phase of intrauterine development. Material and methods: The human conceptus tissue was collected from the Department of Gynecology and Obstetrics and the Department of Pathology after tubal pregnancies or after spontaneous abortions. The immunohistochemistry procedure was used to visualize the targeted proteins. The data acquisition and analysis were performed by immunofluorescence microscopy and captured by a cooled digital camera. The further processing of the pictures was done by Image J and Photoshop Adobe. Results: Panx1 is expressed in all areas of interest during the embryonal development of human spinal cord and surrounding structures and can relate to presence of Panx1 in neuronal tissue, as it was partially co-localized with PGP9.5. There is a change in structural arrangement during the different ages of intrauterine development. Furthermore, the Panx1 expression was also found in ganglia, including dorsal root ganglia (DRG), intramural ganglia, pre-, and paravertebral ganglia. Nevertheless, Panx1 expression was most prominent in DRG, in comparison to the other neural structures studied and were stronger in more developed specimens. Additionally, we investigated the co-localization with NF200, CD31 and Ki67 in the 7th DW. We found rare expression of NF200 in BP of SC and rare co-localization with Panx1. The developing vascular structures of the spinal cord and developing meninges express Panx1 immunoreactivity. There was a complex pattern, showing the expression of Panx1 in endothelial cells and CD31-non-reactive cells in vascular wall. The co-localization of Ki67 with Panx1 was demonstrated in meninges, notochord, inner layer, and developing blood vessels. Conclusion: The spatiotemporal expression pattern of Panx1 during early intrauterine development of human SC and ganglia was characterized. The presence of Panx1 immunoreactivity supports its possible role in processes of neuronal formation, and migration, in the SC and ganglia during development. These data might help us to understand potential pathology during development of SC and peripheral nervous system.
Ciljevi: Cilj ove studije bio je istražiti izražaj paneksina 1 (Panx1) u kralježničnoj moždini i ganglijima ljudskih zametaka tijekom rane faze intrauterinog razvoja. Materijali i metode: Tkivo ljudskih konceptusa prikupljeno je iz Zavoda za ginekologiju i opstetriciju i Zavodu za patologiju nakon spontanih pobačaja ili nakon tubarne trudnoće. Za vizualizaciju ciljanih proteina korišten je imunohistokemijski postupak. Prikupljanje i analiza podataka provedeni su imunofluorescencijskom mikroskopijom, a snimanje obavljeno hlađenom digitalnom kamerom. Daljnja obrada slika izvršena je korištenjem Image J i Adobe Photoshop softvera. Rezultati: Panx1 je bio izražen u svim područjima od interesa tijekom embrionalnog razvoja ljudske kralježnične moždine i okolnih struktura, što se može povezati s prisutnošću Panx1 u živčanom tkivu, budući da je bio djelomično ko-lokaliziran s PGP9.5. Postoji promjena u strukturnom rasporedu tijekom različitih razdoblja intrauterinog razvoja. Nadalje, izražaj Panx1 također je pronađena u ganglijima, uključujući spinalne ganglije, intramuralne ganglije, pre- i paravertebralne ganglije. Unatoč tome, ekspresija Panx1 bila je najizraženija u spinalnom gangliju, u usporedbi s drugim proučavanim neuralnim strukturama i bila je jača u razvijenijih uzoraka. Osim toga, istražili smo ko-lokalizaciju s NF200, CD31 i Ki67 u 7. DW. Vrlo iznimno pronašli smo izražaj NF200 u bazalnoj ploči (BP) kralježnične moždine, koji je samo rijetko ko-lokalizirao s Panx1. Vaskularne strukture leđne moždine u razvoju i moždane ovojnice u razvoju izražavaju Panx1 imunoreaktivnost. Postojao je složeni uzorak, koji pokazuje ekspresiju Panx1 u endotelnim stanicama i CD31-nereaktivnim stanicama u vaskularnoj stijenci. Ko-lokalizacija Ki67 s Panx1 dokazana je u moždanim ovojnicama, notokordu, unutarnjem sloju i krvnim žilama u razvoju. Zaključci: Okarakteriziran je prostorno-vremenski izražaj Panx1 tijekom ranog intrauterinog razvoja kralježnjićne moždine i ganglija čovjeka. Prisutnost Panx1 imunoreaktivnosti ukazuje na njegovu moguću ulogu u procesima formiranja i migracije neurona u kralježničnoj moždini i ganglijima tijekom razvoja. Ovi podaci mogu pomoći u razumijevanju potencijalne patologije tijekom razvoja SC i perifernog živčanog sustava