Effect Of β-1,3-Galactosyltransferase 2 Gene Knockout On Neurons,Glia Cells And Delta Opioid Receptor In Trigeminal Ganglia Of Adult Mice

ObjectiveTrigeminal ganglion(TG)is the main primary afferent neuron in the trigeminal nerve conduction pathway,which is mainly responsible for regulating various sensory functions of the maxillofacial region.Delta opioid receptor(DOR)is expressed in the cell body and nerve fibers of trigeminal ganglion neurons,and plays an important role in the regulation of pain,inflammation and nerve injury related to the trigeminal nervous system.Glycosyltransferase is a key enzyme catalyzing the formation of a variety of glycosylated products with multiple biological functions.There are many types of glycosyltransferase,including β-1,3-galactosyltransferase 2(B3galt2),a member of the β-1,3-galactosyltransferase family,is highly expressed in the central and peripheral nervous systems.Published results have shown that B3galt2 is expressed in trigeminal ganglion and plays an important role in the sensitization of neurons in trigeminal ganglion induced by peripheral nerve inflammation.However,the role of β-1,3-galactosyltransferase 2 gene in normal trigeminal ganglion is not well understood.In this study,β-1,3-galactosyltransferase 2 gene knockout mice were used to investigate the role of B3galt2 in trigeminal ganglion of adult mice and its mechanism,which will provide a necessary theoretical basis for further study of the role of the trigeminal nervous system and related diseases.MethodsHeterozygous mice with β-1,3-galactosyltransferase 2 gene knockout were bred and genotyped by PCR using genomic DNA to obtain adult(6 months)wild-type heterozygous and homozygous mice of the same sex from the same brood.The expression of β-1,3-galactosyltransferase in trigeminal ganglion was observed by X-gal staining and β-gal immunofluorescence technique.The changes of neurons and sensory nerves in trigeminal ganglion of wild-type heterozygous and homozygous mice were compared using anti-Neu N and anti-Peripherin antibodies via immunofluorescence technique.At the same time,anti-GFAP and anti-Iba1 antibodies were used to observe the changes of glial cells in trigeminal ganglia of wild-type heterozygous and homozygous mice,respectively.In addition,immunofluorescence and real-time quantitative PCR were also used to observe and analyze the expression levels of Delta opioid receptor proteins and genes in trigeminal ganglion wild-type of heterozygous and homozygous mice.Results1 、 Adult wild-type heterozygous and homozygous mice from the same brood and sex were successfully bred in the cage with heterozygous β-1,3-galactosyltransferase 2 gene knockout mice.The expression of β-1,3-galactosyltransferase 2 gene in trigeminal ganglia of adult mice was observed by X-gal staining.2、β-1,3-galactosyltransferase 2 gene knockout significantly decreased the number of mature neurons and sensory neurons in the trigeminal ganglion of adult mice and significantly increased the expression of glial cell marker proteins(GFAP and Iba1).Compared with heterozygous mice,the changes of neurons and glial cells in the trigeminal ganglion of homozygous mice were more obvious.3、β-1,3-galactosyltransferase 2 gene knockout significantly decreased the protein and gene expression of Delta opioid receptor in trigeminal ganglion of adult mice.ConclusionsThe β-1,3-galactosyltransferase 2 gene knockout affects neurons,glial cells and Delta opioid receptors in the trigeminal ganglion of adult mice,suggesting that β-1,3-galactosyltransferase 2 gene plays an important role in the trigeminal ganglion.This study provides a certain experimental basis for further study on the role and mechanism of β-1,3-galactosyltransferase 2 in trigeminal neuralgia.