Expression of interneurons in the striatum of genetically modified mouse B4Galnt1.

Uvod: Živčani sustav ljudi građen je od neurona koji tvore neuronske krugove u kojima sudjeluju i interneuroni. Oni omogućuju komunikaciju između središnjega živčanog sustava (SŽS) i motoričkih ili osjetilnih neurona te eksprimiraju proteine koji vežu kalcij kao što su parvalbumin, kalbindin te kalretinin. Gangliozidi su glikosfingolipidi koji se nalaze u svim tkivima, ali ih ponajviše ima u mozgu. Stupaju u interakcije sa receptorima s ciljem reguliranja proliferacije, apoptoze te diferencijacije. U staničnoj jezgri sudjeluju u ravnoteži iona kalcija. Nedostatak u sintezi gangliozida i interneurona dovodi do neuroloških poremećaja. Ciljevi: Odrediti ekspresiju interneurona u striatumu genetički izmjenjenoga miša B4Galnt1 (KO) i miša divljeg tipa (WT) koristeći protutijela za parvalbuminske, kalbindinske i kalretininske interneurone te odrediti postoji li razlika u ekspresiji interneurona u navedenih miševa. Materijali i metode: U istraživanju su korišteni uzorci moždanoga tkiva genetički izmijenjenoga miša i divljega tipa miša. Nakon utvrđivanja genotipa, životinje su žrtvovane, a mozgovi su rezani na kriostatu. Imunohistokemijska analiza provedena je koristeći protutijela za parvalbuminske, kalbindinske i kalretininske interneurone, a dobiveni rezultati analizirani su u računalnim programima ImageJ i Statistica 12. Rezultati: Ekspresija interneurona u striatumu KO miša smanjena je u odnosu na ekspresiju interneurona WT. Istraživanjem je utvrđeno da postoji statistički značajna razlika u ekspresiji kalbindina, ali ne i za parvalbumin i kalretinin. Zaključak: Takvi rezultati podupiru teoriju da gangliozidi služe kao modulatori proteina koji vežu kalcij te utječu na vezanje liganda. Poremećaji u sintezi gangliozida dovode do smanjene ekspresije interneurona u striatumu.Introduction: The human nervous system consists of neurons that build neuron circuits in which interneurons also take part. They enable the communication between the central nervous system and the motor or sensor neurons. Also, they express proteins that bind calcium such as parvalbumin, calretinin and calbindin. Gangliosides are glycosphingolipids that can be found in tissues, but the biggest number of them is in the brain. They interact with receptors with the aim of regulating proliferation, apoptosis and differentiation. In the cell nucleus, they take part in balancing calcium ions. Deficiency in the synthesis of gangliosides and interneurons leads to neurological disorders. Goals: The aim of this study was to determine expression of interneurons in striatum of a genetically modified mouse B4Galnt1 (KO) and a wild type (WT) by using antibodies for parvalbumin, calbindin and calretinin interneurons, and to determine if there is a difference in the expression of interneurons between a wild type mouse and a genetically modified mouse. Materials and methods: In this research, brain tissue samples from a genetically modified mouse and a wild type of mouse were used. After determining the genotype, the animals were done away with, and the brains were cut on cryostat. An immunohistochemical analysis was carried out by using specific antibodies for parvalbumin, calbindin and calretinin interneuorns, and the results were analysed in two softwares: ImageJ and Statistica 12. Results: The expression of interneurons in the striatum of the genetically modified mouse was reduced in comparison with the expression of interneurons in the wild type mouse. The research confirmed that there is a statistically significant difference in expression of calbindin, but not parvalbumin and calretinin. Conclusion: These results confirm the theory that gangliosides serve as protein modulators that bind calcium and impact binding ligand properties. The disorders in the synthesis of gangliosides lead to a reduced expression of interneurons in the striatum